Cytomics FC 500 - Rutgers...

Cytomics FC 500With CXP Software

Instructions For Use

PN 624923B (June 2004)

Beckman Coulter, Inc.Fullerton, CA 92835

WARNINGS AND PRECAUTIONS

R E AD AL L PR O D U CT M ANU AL S AND CO NS U L T W IT H BE CK M AN CO U L T E R -T R AINE D PE R S O NNE L BE FO R E AT T E M PT ING T O O PE R AT E INS T R U M E NT . D O NO T AT T E M PT T O PE R FO R M ANY PR O CE D U R E BE FO R E CAR E FU L L Y R E AD ING AL L INS T R U CT IO NS . AL W AY S FO L L O W PR O D U CT L ABE L ING AND M ANU FACT U R E R ’S R E CO M M E ND AT IO NS . IF IN D O U BT AS T O H O W T O PR O CE E D IN ANY S IT U AT IO N, CO NT ACT Y O U R BE CK M AN CO U L T E R R E PR E S E NT AT IV E .

BE CK M AN CO U L T E R , INC. U R G E S IT S CU S T O M E R S T O CO M PL Y W IT H AL L NAT IO NAL H E AL T H AND S AFE T Y S T AND AR D S S U CH AS T H E U S E O F BAR R IE R PR O T E CT IO N. T H IS M AY INCL U D E , BU T IT IS NO T L IM IT E D T O , PR O T E CT IV E E Y E W E AR , G L O V E S , AND S U IT ABL E L ABO R AT O R Y AT T IR E W H E N O PE R AT ING O R M AINT AINING T H IS O R ANY O T H E R AU T O M AT E D L ABO R AT O R Y ANAL Y Z E R .

HAZ ARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS

W AR NING S , CAU T IO NS , and IM PO R T ANT S alert y ou as follow s :

WARNING - Can caus e injury .

CAUTION - Can caus e d amag e to th e ins trument.

IMPORTANT - Can caus e mis lead ing res ults .

WARNING R is k of op erator injury if:

r All d oors , covers and p anels are not clos ed and s ecured in p lace p rior to and d uring ins trument op eration.

r T h e integ rity of s afety interlocks and s ens ors is comp romis ed .

r Ins trument alarms and error mes s ag es are not acknow led g ed and acted up on.

r Y ou contact moving p arts .

r Y ou mis h and le b roken p arts .

r D oors , covers and p anels are not op ened , clos ed , removed and /or rep laced w ith care.

r Imp rop er tools are us ed for troub les h ooting .

T o avoid injury :

r K eep d oors , covers and p anels clos ed and s ecured in p lace w h ile th e ins trument is in us e.

r T ake full ad vantag e of th e s afety features of th e ins trument. D o not d efeat s afety interlocks and s ens ors .

r Acknow led g e and act up on ins trument alarms and error mes s ag es .

r K eep aw ay from moving p arts .

r R ep ort any b roken p arts to y our Beckman Coulter R ep res entative.

r O p en/remove and clos e/rep lace d oors , covers and p anels w ith care.

r U s e th e p rop er tools w h en troub les h ooting .

CAUTION S y s tem integ rity mig h t b e comp romis ed and op erational failures mig h t occur if:

r T h is eq uip ment is us ed in a manner oth er th an s p ecified . O p erate th e ins trument as ins tructed in th e Prod uct M anuals .

r Y ou introd uce s oftw are th at is not auth oriz ed b y Beckman Coulter into y our comp uter. O nly op erate y our s y s tem’s comp uter w ith s oftw are auth oriz ed b y Beckman Coulter.

r Y ou ins tall s oftw are th at is not an orig inal cop y rig h ted vers ion. O nly us e s oftw are th at is an orig inal cop y rig h ted vers ion to p revent virus contamination.

IMPORTANT If y ou p urch as ed th is p rod uct from any one oth er th an Beckman Coulter or an auth oriz ed Beckman Coulter d is trib utor, and , if it is not p res ently und er a Beckman Coulter s ervice maintenance ag reement, Beckman Coulter cannot g uarantee th at th e p rod uct is fitted w ith th e mos t current mand atory eng ineering revis ions or th at y ou w ill receive th e mos t current information b ulletins concerning th e p rod uct. If y ou p urch as ed th is p rod uct from a th ird p arty and w ould like furth er information concerning th is top ic, call y our Beckman Coulter R ep res entative.

iiiPN 624923B

REVISION STATUS

Issu e A, I n itial Issu e, 8 /03CX P S o ftw a r e V e r s io n 1.0 . Initia l is s u e fo r c u s to m e r dis tr ib u tio n.

Issu e B, 6/04CX P S o ftw a r e V e r s io n 2.0 .

A dditio ns , c h a ng e s o r de le tio ns w e r e m a de a t th e s e lo c a tio ns :1-3, 1-5, 1-31, 1-32, 2-1, 2-8 , 2-9, 2-11, 3-1, 3-6, 3-11, 3-13, 4-4, 4-5, 4-7 , 4-9, 4-10 , 4-12, 5-7 , 5-11, 5-15 & 6-10 .

This document applies to the latest software listed and higher versions. When a subsequent software version changes the information in this document, a new issue will be released.

PN 624923Biv

REVISION STATUS

PN 624923B v

CONTENTS

WAR N I N G S AN D PR E CAU TIO N S

R E V ISIO N STATU S, iii

I N TR O D U CTIO N , x iii

U S IN G Y O U R Cy to m ic s F C 50 0 MA N U A LS , x iii

A B O U T T HIS MA N U A L, x iii

CO N V E N T IO N S , x ivT o Ch o o s e A Co m m a nd W ith T h e K e y b o a rd, x vD ia lo g B o x , x vD e s c r ip tio n o f R e p o r ting U nits , x v

G R A P HICS , x v

1 SY STE M O V E R V I E W, 1-1

1.1 IN T E N D E D U S E , 1-1A p p lic a tio ns fo r th e Ins tr u m e nt, 1-2

1.2 CY T O ME T E R CO N T R O LS A N D IN D ICA T O R S , 1-3F C 50 0 F lo w Cy to m e tr y S y s te m , 1-3Cy to m e te r Indic a to r P a ne l, 1-4Cy to m e te r R e a dy a nd La s e r O n Indic a to r s , 1-4S a m p le F lo w R a te Indic a to r s , 1-5S ig na l A m p litu de Indic a to r s , 1-5Le v e l S e ns e a nd F lo w Indic a to r s , 1-6MCL ( Mu lti-tu b e Ca r o u s e l Lo a de r ) , 1-6W o r k s ta tio n, 1-7P o w e r S u p p ly , 1-7P r inte r ( O p tio na l) , 1-7

1.3 MICR O S O F T ® W IN D O W S ® D E S K T O P , 1-8W indo w s 20 0 0 A dm inis tr a to r P a s s w o rd, 1-8T a s k b a r, 1-8S ta r t B u tto n, 1-8R e c y c le B in, 1-9My Co m p u te r, 1-10Co ntr o l P a ne l, 1-10U s ing a Mo u s e , 1-10Co lo r R e s o lu tio n, 1-11

1.4 LE A R N I N G T HE B A S IC O P E R A T IN G T E CHN IQ U E S , 1-13MCL Ca r o u s e ls , 1-13B a r-Co de La b e ls , 1-13P u tting a B a r-Co de La b e l o n a S a m p le T u b e , 1-13P u tting S a m p le T u b e s in a Ca r o u s e l, 1-14P u tting a Ca r o u s e l in th e MCL, 1-14R e m o v ing a Ca r o u s e l fr o m th e MCL, 1-16

PN 624923Bvi

CONTENTS

1.5 CX P S O F T W A R E S HO R T CU T S , 1-17K e y b o a rd S h o r tc u ts , 1-17D r a g A nd D r o p , 1-18T o o lb a r B u tto ns , 1-19

1.6 CR E A T IN G P R O T O CO LS , 1-20

1.7 CR E A T IN G R E G I O N S , 1-22Cre a te P o ly g o na l R e g io ns , 1-22Cre a te R e c ta ng u la r R e g io ns , 1-23Cre a te Q u a dr a nt R e g io ns , 1-24

R e p o s itio ning a Q u a dr a nt R e g io n, 1-25Cre a te Line a r R e g io ns , 1-26Cre a te Mu ltip le Line a r R e g io ns , 1-27

1.8 CR E A T IN G G A T E S , 1-28Cre a te A u to G a te , 1-28

Co nv e r t a P o ly g o na l R e g io n to a n A u to G a te R e g io n, 1-30A u to m a tic G a te Cre a tio n, 1-30

1.9 CR E A T IN G F LO W P A G E S , 1-31F lo w P A G E E x a m p le , 1-31D e le te F lo w P A G E S , 1-32

1.10 CR E A T IN G P A N E LS , 1-32U s e P lo ts a nd G a te s fr o m P r e v io u s T e s t, 1-34U s e P lo ts a nd G a te s fr o m P r o to c o l, 1-34U s e Ins tr u m e nt S e tting s , 1-34U s e R e g io ns fr o m P r o to c o l, 1-34Ca r r y R e g io ns fr o m P r e v io u s P r o to c o l, 1-34U s e R e g io ns fr o m T h is P r o to c o l:, 1-34U s e Ins tr u m e nt S e tting s fr o m P r e v io u s T e s t, 1-34U s e Ins tr u m e nt S e tting s fr o m E x te rna l F ile , 1-34P a r a m e te r N a m e s , 1-34

1.11 CR E A T IN G W O R K LIS T S , 1-35W o r k lis t P a ne l, 1-35W o r k lis t T e s t, 1-36

1.12 U S IN G T HE O N LIN E HE LP S Y S T E M, 1-37A c c e s s O nline He lp , 1-37Ho w to U s e He lp , 1-37He lp Me nu , 1-38

CX P He lp , 1-38A b o u t... , 1-38O nline S u p p o r t, 1-38

1.13 P E R F O R MA N CE CHA R A CT E R I S T ICS , 1-38

2 D AIL Y R O U TIN E , 2-1

2.1 B E F O R E Y O U B E G IN , 2-1

PN 624923B vii

CONTENTS

2.2 D A ILY S T A R T U P , 2-1Ch e c k W a s te a nd R e a g e nt Le v e ls , 2-2P o w e r th e Co m p u te r a nd Cy to m e te r O N , 2-3

T u rning O n P o w e r, 2-3Lo g g ing O nto W indo w s S o ftw a r e , 2-4Lo g g ing O nto CX P S o ftw a r e , 2-4

Ch e c k th e P o w e r S u p p ly , 2-6A dditio na l S ta r t U p Ch e c k s , 2-8

2.3 D A ILY S HU T D O W N , 2-9W h e n to S h u t D o w n th e Cy to m e te r, 2-9B e fo r e P e r fo r m ing S h u t D o w n, 2-9P o w e r th e Co m p u te r a nd Cy to m e te r O F F , 2-9A fte r Ins tr u m e nt S h u t D o w n, 2-11

2.4 E X T E N D E D S HU T D O W N , 2-12

2.5 CY T O ME T E R A U T O S T A R T U P , 2-14

3 Q U AL ITY CO N TR O L , 3-1

3.1 IN T R O D U CT IO N , 3-1

3.2 Q C P R O CE S S E S , 3-1

3.3 D A ILY Q C, 3-2P r e p a r ing A u to S e tu p S a m p le s , 3-2

3.4 A U T O S E T U P S CHE D U LE R , 3-4

3.5 A U T O S E T U P I I W I Z A R D , 3-5B e fo r e R u nning th e A u to S e tu p I I W iz a rd, 3-5

3.6 A P P LICA T IO N D E F I N IT I O N W I Z A R D , 3-10B e fo r e R u nning th e A p p lic a tio n D e finitio n W iz a rd, 3-10S e tting U p Q C P r o du c ts , 3-14E x p o r ting R e g io n R e s u lts T o Q C D a ta b a s e a nd A s s ig ning Q C P r o du c ts , 3-14R e v ie w Q C R e s u lts , 3-15

4 R U N N I N G SAM PL E S, 4-1

4.1 S A MP LE R E Q U IR E ME N T S , 4-1

4.2 B E F O R E R U N N IN G S A MP LE S , 4-1

4.3 R U N N IN G S A MP LE S - MCL A U T O MA T IC MO D E , 4-2

4.4 R U N N IN G S A MP LE S - MCL MA N U A L MO D E , 4-5

4.5 R U N N IN G S A MP LE S - S IN G LE T U B E MO D E , 4-9

5 O PE R ATIO N PR IN CIPL E S , 5-1

5.1 W HA T T HIS CHA P T E R E X P LA IN S , 5-1

PN 624923Bviii

CONTENTS

5.2 S A MP LE F LO W , 5-1S a m p le Lo a ding , 5-1Hy dro dy na m ic F o c u s ing , 5-1

5.3 LA S E R B E A M S HA P IN G , 5-3

5.4 CE LL ILLU MIN A T IO N , 5-4F o r w a rd S c a tte r, 5-4S ide S c a tte r a nd F lu o r e s c e nt Lig h t, 5-4

5.5 LIG HT CO LLE CT IO N , S E P A R A T I O N A N D ME A S U R E ME N T , 5-4F o r w a rd S c a tte r Co lle c tio n, 5-4S ide S c a tte r a nd F lu o r e s c e nt Lig h t Co lle c tio n, 5-6

S ide S c a tte r, 5-6F lu o r e s c e nt Lig h t, 5-7

5.6 S IG N A L P R O CE S S IN G , 5-8V o lta g e P u ls e S ig na ls , 5-8

P e a k S ig na l, 5-8Inte g r a l S ig na l, 5-10

A m p lific a tio n, 5-10S ig na ls G e ne r a te d, 5-10

5.7 P R O T O CO LS , 5-11 S p e c ia l P r o to c o ls a nd P a ne ls , 5-11

S p e c ia l P a ne ls , 5-11

5.8 A U T O MA T E D S O F T W A R E F E A T U R E S , 5-12

5.9 P A R A ME T E R S , 5-12A U X ilia r y P a r a m e te r, 5-12

W h e n to U s e th e A U X P a r a m e te r, 5-12T IME P a r a m e te r, 5-12R A T IO P a r a m e te r, 5-13

5.10 P LO T D IS P LA Y , 5-13P R IS M, 5-13R e g io ns , 5-15G a ting , 5-15D a ta S to r a g e , 5-15His to g r a m S ta tis tic s , 5-16

Line a r R e g io n S ta tis tic s , 5-16Lo g R e g io n S ta tis tic s , 5-17Lo g -Lo g Me a n Me th o d, 5-17Lin-Lo g Me a n Me th o d, 5-18

6 TR O U BL E SH O O TIN G , 6-1

6.1 P R E CA U T I O N S /HA Z A R D S , 6-1La s e r /R a dia tio n P r e c a u tio ns , 6-1La s e r W a rning La b e ls , 6-1W a rning La b e ls o n U P S , 6-10

PN 624923B ix

CONTENTS

D is p o s a l P r e c a u tio n, 6-11

6.2 MA IN T E N A N CE S CHE D U LE S , 6-11Cle a ning S c h e du le , 6-11R e p la c e m e nt S c h e du le , 6-11

6.3 E R R O R ME S S A G E S , 6-11D is p la y Lo c a tio ns , 6-11

CX P S o ftw a r e S c r e e n, 6-11Cy to m e te r S ta tu s Me s s a g e s S c r e e n, 6-12Cy to m e te r.lo g F ile , 6-12

6.4 E R R O R ME S S A G E CY T O ME T E R .LO G F ILE , 6-13Ho w to A c c e s s th e Cy to m e te r.lo g F ile , 6-13

F r o m CX P S o ftw a r e , 6-13F r o m W indo w s D e s k to p , 6-13

Cy to m e te r.Lo g E ntr y D e s c r ip tio n, 6-14Ho w to S e a r c h th e Cy to m e te r.lo g F ile , 6-14O th e r F u nc tio ns A v a ila b le , 6-14

P r int, 6-14Ch a ng e F o nt, 6-14F ind He lp T o p ic s , 6-14

Cy to m e te r a rc h iv e .lo g F ile , 6-14

6.5 E R R O R ME S S A G E S T A B LE , 6-15

6.6 LE V E L S E N S E IN D ICA T O R S , 6-20S h e a th Lo w , 6-20W a s te F u ll, 6-20

6.7 CY T O ME T E R CO N T R O L W IN D O W CA N N O T B E V I E W E D , 6-20

A H O W TO ..., A-1

A .1 S O F T W A R E P R O CE D U R E S , A -1T h e s e p r o c e du r e s a r e in th e S y s te m O v e r v ie w c h a p te r in th e Ins tr u c tio ns F o r U s e Ma nu a l., A -1T h e s e p r o c e du r e s a r e in th e Q u a lity Co ntr o l c h a p te r in th e Ins tr u c tio ns F o r U s e Ma nu a l., A -1T h e s e p r o c e du r e s a r e in th e U s ing CX P S o ftw a r e c h a p te r in th e R e fe r e nc e Ma nu a l., A -1

A .2 HA R D W A R E P R O CE D U R E S , A -1T h e s e p r o c e du r e s a r e in th e S p e c ia l P r o c e du r e s Ma nu a l., A -1

B BAR -CO D E SPE CIFICATIO N S, B-1

B .1 B A R -CO D E S A MP LE ID E N T IF ICA T IO N , B -1

B .2 CO R R E CT P LA CE ME N T O F T HE B A R -CO D E LA B E L, B -1

B .3 B A R -CO D E LA B E L S P E CIF ICA T IO N S , B -2La b e l S iz e a nd T h ic k ne s s , B -2S y m b o l D im e ns io ns , B -3La b e l a nd P r int Q u a lity , B -3

PN 624923Bx

CONTENTS

B .4 B A R -CO D E E R R O R R A T E , B -4

B .5 B A R -CO D E S Y MB O LO G IE S , B -5

B .6 B A R -CO D E LA B LE S , B -6B a r-Co de La b e l O p tic a l Ch a r a c te r is tic s a t 67 0 nm ± 10 % , B -6

B .7 MCL B A R -CO D E R E A D E R , B -6

B .8 B A R -CO D E D E CO D E R , B -7

B .9 CHE CK S U M A LG O R IT HM, B -7

R E FE R E N CE S, R E FE R E N CE S-1

IN D E X, IN D E X-1

TR AD E M AR K S

PN 624923B xi

ILLUSTRATIONS5.1 F lo w Ce ll ( Hy dr o dy na m ic F o c u s ing ) , 5-25.2 La s e r B e a m S h a p ing , 5-35.3 O p tic a l S y s te m w ith O p tio na l R e d S o lid-s ta te La s e r, 5-55.4 O p tic a l S y s te m w ith O p tio na l He N e La s e r , 5-55.5 S ing le La s e r F ilte r B lo c k Co nfig u r a tio n, 5-65.6 D u a l La s e r F ilte r B lo c k Co nfig u r a tio n, 5-75.7 V o lta g e P u ls e F o r m a tio n, P e a k S ig na l, 5-95.8 Inte g r a l a nd P e a k P u ls e s , 5-105.9 P r is m P lo t, 5-146.1 La s e r La b e ls o n th e S e ns ing Co m p a r tm e nt Co v e r, 6-26.2 La s e r La b e ls in th e O p tic a l A r e a , F r o nt V ie w , 6-36.3 La s e r La b e ls in th e O p tic a l A r e a , Inte r io r V ie w , 6-46.4 La s e r La b e ls o n th e A r g o n La s e r He a d, 6-56.5 La s e r La b e ls fo r O p tio na l R e d S o lid-s ta te La s e r, 6-56.6 Lo c a tio n o f th e O p tio na l R e d S o lid-s ta te La s e r in th e Ins tr u m e nt, R e a r V ie w , 6-66.7 La s e r La b e ls fo r O p tio na l R e d He liu m - N e o n La s e r, 6-66.8 Lo c a tio n o f th e O p tio na l R e d He liu m -N e o n La s e r in th e Ins tr u m e nt, R e a r V ie w , 6-76.9 La s e r La b e ls o n th e Cy to m e te r B a c k P a ne l, 6-76.10 La s e r La b e ls o n th e MCL P r o b e Ho u s ing Co v e r, 6-86.11 La s e r La b e ls o n th e MCL B a r-Co de R e a de r, 6-96.12 Inte rna tio na l W a rning S y m b o l Lo c a tio ns , 6-106.13 E r r o r Me s s a g e o n CX P S o ftw a r e S c r e e n, E x a m p le , 6-126.14 Cy to m e te r S ta tu s Me s s a g e s W indo w , E x a m p le , 6-126.15 E x a m p le o f E r r o r Me s s a g e s in Cy to m e te r.lo g F ile , 6-136.16 S h e a th Lo w Indic a to r s , 6-206.17 W a s te F u ll Indic a to r s , 6-20B .1 B a r-Co de La b e l, B -1B .2 B a r-Co de La b e l P la c e m e nt, B -2B .3 B a r-Co de La b e l S p e c ific a tio ns , B -3

PN 624923B xii

TABLES1.1 A p p lic a tio ns fo r th e Ins tr u m e nt, 1-26.1 Cle a ning S c h e du le , 6-116.2 E r r o r Me s s a g e s , 6-15B .1 B a r-Co de S y m b o lo g ie s , B -5B .2 Co de -R e la te d S p e c ific a tio ns , B -6

PN 624923B xiii

INTRODUCTION

T h is intr o du c to r y s e c tio n c o nta ins th e fo llo w ing to p ic s :

r U S I N G Y O U R Cy to m ic s F C 50 0 MA N U A LS

r A B O U T T HIS MA N U A L

r CO N V E N T IO N S , a nd

r G R A P HICS .

USING Y OUR Cy tom ics FC 50 0 MANUALST h e m a nu a ls lis te d b e lo w a r e a v a ila b le a s P D F file s in th e F C 50 0 CX P s o ftw a r e a nd th e O p e r a to r ’s Ma nu a ls CD -R O M. P r inte d v e r s io ns o f th e s e m a nu a ls a r e a ls o a v a ila b le b y o rde r.

U s e th e R eferen ce m a nu a l fo r ins tr u m e nt s p e c ific a tio ns a nd info r m a tio n o n ins ta lla tio n a nd s y s te m o p tio ns .

U s e th e I n stru ction s For U se m a nu a l fo r th e da y -to -da y r u nning o f y o u r ins tr u m e nt. G o th r o u g h th e de ta ile d s te p -b y -s te p p r o c e du r e s o f s ta r tu p , q u a lity c o ntr o l ( Q C) , r u nning s a m p le s , a na ly z ing da ta , p r inting r e p o r ts , r e v ie w ing Q C da ta , s h u tdo w n. It c o nta ins s a fe ty a nd tr o u b le s h o o ting info r m a tio n, e r r o r m e s s a g e s , a s w e ll a s in-de p th info r m a tio n o n th e p r inc ip le s o f flo w c y to m e tr y , info r m a tio n a b o u t w h a t y o u r ins tr u m e nt do e s , a nd th e m e th o ds it u s e s .

U s e th e Sp ecial Proced u res m a nu a l to c le a n, r e p la c e , o r a dju s t a c o m p o ne nt o f th e ins tr u m e nt.

U s e th e G ettin g Started m a nu a l a s a b r ie f intr o du c tio n to th e s y s te m .

U s e th e M aster I n d ex to e a s ily lo c a te a to p ic in a ny o f y o u r m a nu a ls .

ABOUT THIS MANUALT h e info r m a tio n in y o u r F C 50 0 Ins tr u c tio ns F o r U s e m a nu a l is o r g a niz e d a s fo llo w s :

r Ch a p te r 1 S Y S T E M O V E R V IE WP r o v ide s th e inte nde d u s e o f th e ins tr u m e nt, th e c o ntr o ls a nd indic a to r s , a nd info r m a tio n o n u s ing th e s y s te m ’s s o ftw a r e a nd W o r k s ta tio n.

r Ch a p te r 2 D A ILY R O U T IN EP r o v ide s info r m a tio n o n do ing da ily p r o c e du r e s , s u c h a s S ta r tu p a nd S h u tdo w n.

r Ch a p te r 3 Q U A LIT Y CO N T R O L P r o v ide s info r m a tio n o n h o w to r u n q u a lity c o ntr o l m a te r ia l to v e r ify ins tr u m e nt s e tu p .

r Ch a p te r 4 R U N N IN G S A MP LE SP r o v ide s info r m a tio n o n h o w to r u n p a tie nt b lo o d s a m p le s .

s Ch a p te r 5, O P E R A T I O N P R I N CIP LE SCo nta ins a de s c r ip tio n o f flo w c y to m e tr y , th e no r m a l s a m p le flo w th r o u g h th e ins tr u m e nt, h o w lig h t c o lle c tio n a nd s ig na l p r o c e s s ing a r e a c c o m p lis h e d a nd h o w th e p a r a m e te r s a r e de r iv e d.

r Ch a p te r 6, T R O U B LE S HO O T IN G

D e s c r ib e s la s e r s a fe ty p r e c a u tio ns a nd th e lo c a tio n o f th e la s e r- r e la te d la b e ls a nd h o w to tr o u b le s h o o t th e s y s te m w ith th e e r r o r m e s s a g e s th a t a p p e a r o n th e s c r e e n.

PN 624923Bxiv

INTRODUCTIONCONVENTIONS

s A p p e ndic e sT h e a p p e ndic e s p r o v ide r e fe r e nc e m a te r ia l o n th e fo llo w ing to p ic s :

r HO W T O ...

r B A R -CO D E S P E CIF ICA T IO N S

r IN D E XP r o v ide s p a g e nu m b e r s fo r inde x e d info r m a tio n.

CONVENTIONST h is m a nu a l u s e s th e fo llo w ing c o nv e ntio ns :

r T h r o u g h o u t th is m a nu a l y o u r F C 50 0 is a ls o r e fe r r e d to a s th e s y s te m o r ins tr u m e nt.

r Bold font indic a te s a s o ftw a r e o p tio n, s u c h a s Cy tom eter.

r Italics font indic a te s s c r e e n te x t dis p la y e d o n th e ins tr u m e nt, s u c h a s P reparing S amples.

r Courier font indic a te s te x t y o u h a v e to ty p e u s ing th e k e y b o a rd.

r ë indic a te s a k e y ( s u c h a s Û) .

r ë+ë indic a te s th a t th e tw o k e y s lis te d ( s u c h a s Þ+Ê) a r e link e d fo r a s p e c ific fu nc tio n a nd m u s t b e p r e s s e d in th is s e q u e nc e :

a . P r e s s do w n o n th e fir s t k e y lis te d a nd w h ile c o ntinu ing to p r e s s it, p r e s s do w n o n th e s e c o nd k e y lis te d.

b . R e le a s e b o th k e y s a t th e s a m e tim e .

r ë ë indic a te s to p r e s s a nd r e le a s e th e fir s t k e y lis te d th e n p r e s s a nd r e le a s e th e ne x t k e y lis te d. F o r e x a m p le : Y Û.

r Ic o ns /b u tto ns to s e le c t fu nc tio ns o n th e s o ftw a r e s c r e e n a r e s h o w n w ith in te x t.

E x a m p le : .

r indic a te s to u s e th e m o u s e to s e le c t th e s c r e e n b u tto n la b e le d

. S e e U s ing a Mo u s e .

r File tt Sa v e indic a te s to u s e th e m o u s e to s e le c t th e Sa v e ite m o n th e File m e nu .

r É th r o u g h Ô a r e s p e c ia l fu nc tio n k e y s .

r A N ote c o nta ins info r m a tio n th a t is im p o r ta nt to r e m e m b e r o r h e lp fu l in p e r fo r m ing a p r o c e du r e .

r T h e te r m s “ s c r e e n” a nd “ w indo w ” a r e u s e d inte rc h a ng e a b ly .

r SHOW ME m e a ns th e r e is a v ide o a v a ila b le fo r th e p r o c e du r e in th e o nline h e lp .

PN 624923B xv

INTRODUCTIONG R A P H ICS

To Ch oose A Com m a nd With Th e Key b oa rd

A fte r y o u p r e s s Þ, e a c h m e nu na m e h a s o ne le tte r u nde r line d to indic a te w h ic h le tte r to u s e to p u ll do w n th e m e nu . F o r e x a m p le , th e le tte r F in th e F ile m e nu is u nde r line d, p r e s s F to p u ll do w n th e F ile m e nu ; th e le tte r E in th e E dit m e nu is u nde r line d, p r e s s E to p u ll do w n th e E dit m e nu .

Dia log Box

D ia lo g b o x e s r e c e iv e c o m m a nds o r info r m a tio n; fo r e x a m p le , a file na m e dia lo g b o x r e c e iv e s info r m a tio n a b o u t a file na m e .

Descrip tion of Rep orting Units

U nle s s o th e r w is e s ta te d, a ll p a r a m e te r u nits a r e s h o w n in th e U S u nit fo r m a t ( c e lls /µ L) th r o u g h o u t th e m a nu a ls .

GRAPHICSA ll g r a p h ic s , inc lu ding s c r e e ns a nd p r into u ts , a r e fo r illu s tr a tio n p u r p o s e s o nly a nd m u s t no t b e u s e d fo r a ny o th e r p u r p o s e .

Comman d Fu n ction

Û A c c e p ts y o u r s e le c tio n.

È S to p s th e o p e r a tio n, dis c a rding y o u r c h o ic e s .

Ù Mo v e s c u r s o r o v e r diffe r e nt c h o ic e s if th e r e a r e m u ltip le o p tio ns - s e e

W indo w s ® m a nu a ls fo r W indo w s o p e r a tio n v ia k e y b o a rd.

Þ+Ù W h e n y o u h a v e m o r e th a n o ne a p p lic a tio n W indo w o p e n, u s e Þ+Ù to s w itc h b e tw e e n ta s k s .

A c c e p ts th e info r m a tio n y o u h a v e s e le c te d o r ty p e d.

S to p s th e o p e r a tio n, ig no r ing y o u r c h o ic e s .

PN 624923Bxvi

INTRODUCTIONG R A P H ICS

PN 624923B 1 -1

11SY STEM OVERVIEW

T h e info r m a tio n in th is s e c tio n inc lu de s :

r IN T E N D E D U S E

r CY T O ME T E R CO N T R O LS A N D IN D ICA T O R S

r MICR O S O F T ® W IN D O W S ® D E S K T O P

r LE A R N I N G T HE B A S IC O P E R A T IN G T E CHN IQ U E S

r CX P S O F T W A R E S HO R T CU T S

r CR E A T IN G P R O T O CO LS

r CR E A T IN G R E G I O N S

r CR E A T IN G G A T E S

r CR E A T IN G F LO W P A G E S

r CR E A T IN G P A N E LS

r CR E A T IN G W O R K LIS T S

r U S I N G T HE O N LIN E HE LP S Y S T E M

r P E R F O R MA N CE CHA R A CT E R I S T ICS

1.1 INTENDED USET h e Cy to m ic s F C 50 0 is a s y s te m fo r th e q u a lita tiv e a nd q u a ntita tiv e m e a s u r e m e nt o f b io lo g ic a l a nd p h y s ic a l p r o p e r tie s o f c e lls a nd o th e r p a r tic le s . T h e s e p r o p e r tie s a r e m e a s u r e d w h e n th e c e lls p a s s th r o u g h o ne o r tw o la s e r b e a m s in s ing le - file .

T h e ins tr u m e nt c a n s im u lta ne o u s ly m e a s u r e fo r w a rd s c a tte r, s ide s c a tte r, a nd fiv e flu o r e s c e nt dy e s u s ing o ne o r tw o la s e r s a t 48 8 nm a nd e ith e r 635 nm ( S o lid-s ta te la s e r ) o r 633 nm (He N e la s e r ) . T h e r e fo r e , th e ins tr u m e nt c a n p e r fo r m c o r r e la te d m u ltip a r a m e te r a na ly s e s o f indiv idu a l c e lls .

IMPORTANT T h e us e of th e d ata g enerated b y th is ins trument d ep end s up on th e reg ulatory s tatus of th e reag ents y ou us e. If th e reag ent is lab eled b y th e manufacturer “ For R es earch U s e O nly . Not for us e in d iag nos tic p roced ures ,” U S fed eral law p roh ib its th e us e of th e d ata for d iag nos is .

If, h ow ever, th e reag ent s ub s y s tem is lab eled “ For in vitro d iag nos tic us e,” th e entire s y s tem can b e cons id ered a med ical d evice for g enerating d ata to b e us ed for d iag nos is .

PN 624923B1 -2

SY STEM OVERVIEWINTEND ED U SE

Ap p lica tions for th e Instrum ent

T h e F C 50 0 is c o ns ide r e d a n e q u iv a le nt s y s te m to th e CO U LT E R ® E P ICS ® X L™ a nd th e CO U LT E R E P ICS X L-MCL™ flo w c y to m e te r s w h e n u s e d w ith th e fo r w a rd s c a tte r F ie ld S to p in th e fa c to r y s e t p o s itio n ( 1-19° ) .

T a b le 1.1 lis ts th e m a ny a p p lic a tio ns fo r th e ins tr u m e nt. In a dditio n to h u m a n c e lls , o th e r c e ll ty p e s c a n b e a na ly z e d, s u c h a s :

r P la nt c e lls

r Ma r ine p la nk to n

r A nim a l c e lls

r B a c te r ia

Ta b le 1.1 Ap p lica tions for th e Instrum ent

Ap p lica tions Cell Ty p es

Sa m p le Prep a ra tion (refer to th e rea g ent's

p a ck a g e insert) Mea surem ents Pa ra m eters

Cell s urface antig ens

W h ole b lood

Buffy coats

M ononuclear cells

D is s ociated tis s ue

Platelets

Bone M arrow

ImmunoPrep ™ reag ents w ith

th e Q -PR E P™ W orks tation,

th e M ulti-Q -Prep ™

W orks tation or th e T Q -Prep ™ W orks tation

W h ole b lood ly s ing reag ent kit

Cell g rad ient s ep aration

Fluores cent-lab eled antib od ies

T h romb o Fix™ p latelet s tab iliz er

Cell s iz e and g ranularity

FIT C, R D 1 , E CD , PC5, PC7 , APC and APC-CY 7 , PC7

Forw ard and s id e s catter (L og /L inear)

Fluores cence one, tw o, th ree, four and five (L og /L inear)

Pris m

R atio

T ime

AU X /Peak

Nucleic acid s W h ole b lood

D is s ociated tis s ue

Froz en s ections

Paraffin s ections

Bod y fluid s

V arious s taining meth od s , includ ing :

E th id ium b romid e

Prop id ium iod id e

D NA Prep reag ents

L eukaS ure™


G reen, red , and orang e fluores cence

Forw ard and s id e s catter

Fluores cence one, tw o, th ree, four and five (L og /L inear)

R atio, AU X Peak

K inetics W h ole b lood

Buffy coats

M ononuclear cells

T is s ue culture cells

V arious meth od s , includ ing :

Fluo 3 fluores cence


G reen fluores cence

T ime


Fluores cence one

R atio

T ime

PN 624923B 1 -3

SY STEM OVERVIEWCY TOM ETER CONTR OL S A ND IND ICA TOR S 1

1.2 CY TOMETER CONTROLS AND INDICATORS

FC 50 0 Flow Cy tom etry Sy stem

CH

Cell function W h ole b lood

Buffy coats

M ononuclear cells

T is s ue culture cells

V arious meth od s and d y es , includ ing :

D CFH -D A

D iO C5(3)

FD A




Fluores cence one

R eticulocy tes W h ole b lood T h iaz ole orang e corip h os p h ine O (CPO ) Acrid ine O rang e



R ed fluores cence

L og /L inear Forw ard and S id e S catter

L og Fluores cence one

L og Fluores cence four

Ta b le 1.1 Ap p lica tions for th e Instrum ent (Continued)

b

c

d

e

f

g

h

ij 557 1 026B

PN 624923B1 -4

SY STEM OVERVIEWCY TOM ETER CONTR OL S A ND IND ICA TOR S

Cy tom eter Ind ica tor Pa nel

Cy tom eter Rea d y a nd La ser On Ind ica tors

b MCL g Mo u s e

c Cy to m e te r h U S B b o x

d He N e la s e r o p tio n i Co m p u te r

e K e y b o a rd j P o w e r S u p p ly

f Mo nito r

b B lu e m e a ns th e A r g o n la s e r is o n.

c R e d m e a ns th e r e d la s e r s h u tte r is o p e n.

d G r e e n m e a ns Cy to m e te r is o n a nd in th e R e a dy s ta te .

d c b

PN 624923B 1 -5


Sa m p le Flow Ra te Ind ica tors

Sig na l Am p litud e Ind ica tors

N ote: W h e n b o th lo g a nd line a r p a r a m e te r s a r e s e le c te d fo r th e s a m e s e ns o r, th e a m p litu de dis p la y de fa u lts to th e lo g s ig na l.

b Lo w S a m p le F lo w

c Me diu m S a m p le F lo w

d Hig h S a m p le F lo w

b c d

b F L1 e F L4 h R e d m e a ns o v e r r a ng e

c F L2 f F L5 o r A u x i S S

d F L3 g F S j W h ite indic a te s A u x s o u rc e a nd A u x c h a nne l ( F L5)

b c d e f g

hij

PN 624923B1 -6

SY STEM OVERVIEWCY TOM ETER CONTR OL S A ND IND ICA TOR S

Lev el Sense a nd Flow Ind ica tors

MCL (Multi-tub e Ca rousel Loa d er)

b S h e a th F lo w

c W a s te F u ll

d S h e a th Lo w

b c d

PN 624923B 1 -7


Work sta tion

Pow er Sup p ly

Y o u r ins tr u m e nt h a s o ne o f th e s e c o nfig u r a tio ns . F o llo w th e p r o c e du r e s in th e do c u m e nta tio n th a t a p p lie s to y o u r P o w e r S u p p ly c o nfig u r a tio n.

Printer (Op tiona l)

T h is m a nu a l do e s no t e x p la in h o w to u s e th e o p tio na l P r inte r. O p e r a ting ins tr u c tio ns fr o m th e P r inte r m a nu fa c tu r e r a r e inc lu de d w ith th e P r inte r.

NEC MultiSync LCD 1700M+

U n iv ersal Power Su p p ly V oltag e-sp ecific Power Su p p ly

SYSTEM POPWER

MCL POPWER

SYSTEM POPWER

MCL POPWER

WATER TRAP

VAC TRAP

AIR FILTER

VAC

FILTER

SY S VAC

SY S PRESS

30 PSI

PRESSADJ

PN 624923B1 -8

SY STEM OVERVIEWM ICR OSOFT® W IND OW S® D ESK TOP

1.3 MICROSOFT® WINDOWS® DESKTOP

Wind ow s 20 0 0 Ad m inistra tor Pa ssw ord

B e s u r e to m a inta in y o u r W indo w s 20 0 0 A dm inis tr a to r p a s s w o rd in a s e c u r e lo c a tio n. If y o u lo s e o r fo r g e t th e W indo w s 20 0 0 A dm inis tr a to r p a s s w o rd, y o u m u s t r e im a g e y o u r h a rd dr iv e , c a u s ing th e lo s s o f a ll da ta .

Ta sk b a r

A t th e b o tto m o f th e s c r e e n is th e T a s k b a r. It c o nta ins th e S ta r t b u tto n o n th e le ft-h a nd s ide a nd a c lo c k o n th e r ig h t. O th e r ic o ns a r e dis p la y e d o n th e T a s k b a r de p e nding o n th e c o nfig u r a tio n o f th e h a rdw a r e a nd s o ftw a r e o f y o u r p a r tic u la r c o m p u te r.

W h e n a p r o g r a m is o p e ne d, a b u tto n fo r th a t p a r tic u la r p r o g r a m a p p e a r s w ith in th e T a s k b a r.

Y o u c a n s w itc h b e tw e e n p r o g r a m s b y c lic k ing w ith th e m o u s e o n th e s e b u tto ns .

Sta rt Button

T h e S ta r t b u tto n a llo w s y o u to g a in a c c e s s to th e c o m p u te r p r o g r a m s a nd s e tting s . W h e n y o u

c lic k th e m o u s e o n o n th e T a s k b a r, a m e nu a p p e a r s s h o w ing th e c o m p u te r s y s te m ’s m a in p r o g r a m s a nd p r o g r a m g r o u p ic o ns . T h e fo llo w ing CX P s o ftw a r e “ s h o r tc u t” o p tio ns a r e a v a ila b le o n th is D e s k to p Me nu B a r o nly if th e y h a v e b e e n ins ta lle d.

O th e r s ta nda rd W indo w s fu nc tio ns a r e a ls o dis p la y e d a nd th e s e a r e b r ie fly de s c r ib e d b e lo w :

PN 624923B 1 -9

SY STEM OVERVIEWM ICR OSOFT® W IND OW S® D ESK TOP 1

Recy cle Bin

Y o u c a n de le te file s fr o m y o u r c o m p u te r a nd p la c e th e m in th e R e c y c le B in w h e r e y o u c a n, if ne c e s s a r y , r e tr ie v e th e m la te r. Ho w e v e r, if y o u w is h to p e r m a ne ntly r e m o v e file s , th is is a c h ie v e d b y e m p ty ing th e R e c y c le B in.

Prog ra m s: d is p lay s a lis t of p rog rams th at are ins talled on th e comp uter th at y ou can s tart.

Docum ents: lis ts th e mos t recently op ened d ocuments .

Setting s: d is p lay s th e us er-config urab le s y s tem comp onents , includ ing th e Control Panel.

Sea rch : d is p lay s a menu, w h ich allow s y ou to find Files , Fold ers or Comp uters to w h ich y ou are connected if Netw orking .

Help : op ens th e W ind ow s H elp , w h ich p rovid es g uid ance in op erating th e W ind ow s S y s tem.

Run: d is p lay s a s creen d ialog b ox for s tarting or load ing a p rog ram or op ening a file w ith an M S -D O S command .

Sh ut Dow n: d is p lay s a w ind ow d ialog b ox w ith th e op tions for S h utting D ow n th e Comp uter S y s tem, res tarting or oth er s p ecific op tions as d is p lay ed .

PN 624923B1 -1 0


My Com p uter

T h e My Co m p u te r ic o n o n th e W indo w s D e s k to p a llo w s a c c e s s to th e c o nte nts o f y o u r c o m p u te r ( ins ta lle d dr iv e s , th e Co ntr o l P a ne l, P r inte r s a nd o th e r ic o ns a s a p p r o p r ia te to th e w a y th e p a r tic u la r c o m p u te r s y s te m w a s c o nfig u r e d) .

Control Pa nel

T h e Control Pa nel h o lds th e to o ls th a t y o u c a n u s e to a lte r th e w a y W indo w s lo o k s a nd w o r k s .

Using a Mouse

T h e Mo u s e , a h a ndh e ld inp u t de v ic e , c o ntr o ls th e m o v e m e nt o f a p o inte r o n th e c o m p u te r s c r e e n. A s th e m o u s e is m o v e d, th e p o inte r m o v e s o n th e s c r e e n. W h e n y o u p o s itio n th e p o inte r o v e r a n o b je c t o n th e s c r e e n, y o u c a n c lic k th e m o u s e b u tto n to do diffe r e nt ta s k s .

If y ou w is h to s ee th e contents of any d rive or fold er, d oub le click th e mous e on its icon. For M icros oft W ind ow s 2000, th e comp uter h ard d is k is know n as C d rive.

T h e Flop p y D rive is D rive A, and any CD -R O M d rive is know n as D D rive unles s W ind ow s as s ig ns anoth er s uitab le letter.

T o view th e Control Pa nel:

Click on Sta rt and p oint th e mous e to S etting s .

Click on th e Control Panel icon.

Note: T h e icons in th e Control Panel vary accord ing to th e p articular h ard w are and s oftw are s etting s for y our comp uter. W h en y ou click on an icon, a s h ort d es crip tion of th e s etting is s h ow n.

D oub le click on an icon to s ee th e s etting th at y ou can ch ang e for th at item.

For a full d es crip tion of th e p os s ib le s etting s , refer to th e M icros oft W ind ow s H elp text and any b ooks or manuals s up p lied w ith y our comp uter s y s tem.

PN 624923B 1 -1 1

SY STEM OVERVIEWM ICR OSOFT® W IND OW S® D ESK TOP 1

Y o u c a n c lic k th e m o u s e b u tto n to s e le c t file s , do u b le c lic k to o p e n a nd w o r k in file s , o r c lic k a nd dr a g to m o v e file s .

A s m o s t fu nc tio ns inv o lv e p o inting a t a n o b je c t o n th e s c r e e n a nd th e n c lic k ing th e m o u s e b u tto n, th e fo llo w ing s h o w s o m e o f th e c o m m o n a c tio ns :

Y o u c a n s w itc h le ft c a nd r ig h t b m o u s e b u tto ns . F o r m o r e info r m a tio n a b o u t u s ing a m o u s e , s e e W indo w s He lp .

T h e CX P s o ftw a r e is u s e d w ith a m o u s e a nd e x c lu s iv e ly u s e s th e le ft m o u s e b u tto n e x c e p t w h e r e s p e c ific a lly indic a te d w ith in th e He lp T e x t.

Color Resolution

F o r th e b e s t r e s u lts , y o u ne e d to r e s e t y o u r c o m p u te r to a c o lo r r e s o lu tio n o f a t le a s t 10 24 x

7 68 p ix e ls a nd Hig h Co lo r ( 16 b it) .

Click : Pres s and releas e th e left mous e b utton c once.

Select: Point at a p articular ob ject and click once.

Doub le-click : Q uickly p res s and releas e th e left mous e b utton c tw ice.

Rig h t-click : Pres s and releas e th e rig h t mous e b utton b once.

Point: M ove th e mous e until th e curs or is over th e d es ired icon or ob ject.

Press: H old d ow n a mous e b utton.

Dra g : H old d ow n th e left mous e b utton c w h ile moving th e mous e.

b

c

1 Clo s e do w n a ny a p p lic a tio ns th a t y o u m a y h a v e r u nning , inc lu ding a ny th a t h a v e b e e n

m inim iz e d o n th e T o o l B a r.

2 S e le c t My Com p uter a nd th e n c h o o s e th e Co ntr o l P a ne l ic o n.

3 F r o m th e Control Pa nel, c h o o s e th e Disp la y o p tio n.

PN 624923B1 -1 2


N ote: 3dfx v o o do o b a s e d v ide o c a rds m u s t b e s e t to Hig h Co lo r ( 16 b it) fo r 3D a c c e le r a tio n to fu nc tio n c o r r e c tly .

4 S e le c t th e Setting s ta b .

5 Mo v e th e Screen Area s lide r to s e t 10 24

x 7 68 c o lo r r e s o lu tio n.

6 S e t th e Colors dia lo g b o x u s ing th e dr o p

do w n m e nu to s e t Hig h Co lo r ( 16 b it) .

7 to c o m p le te th e

o p e r a tio n. Y o u m a y ne e d to r e s ta r t th e c o m p u te r ( a dia lo g b o x is dis p la y e d if th is is r e q u ir e d) .

PN 624923B 1 -1 3

SY STEM OVERVIEWL EA R NING TH E B A SIC OP ER A TING TECH NIQ U ES 1

1.4 LEARNING THE BASIC OPERATING TECHNIQUESB e fo r e r e a ding th e o th e r c h a p te r s in th is m a nu a l:

r R e a d a b o u t th e Cy to m e te r Co ntr o l s c r e e ns in th e R e fe r e nc e m a nu a l.

r R e a d th is c h a p te r to b e c o m e e x p e r ie nc e d w ith u s ing th e MCL.

P r a c tic e th e b a s ic te c h niq u e s u ntil y o u fe e l c o m fo r ta b le u s ing th e m . If, la te r o n, y o u ne e d to u s e a b a s ic te c h niq u e b u t c a nno t r e m e m b e r h o w , u s e th e O nline He lp Inde x o r S e a rc h ta b to lo o k it u p a nd g e t th e s te p - b y - s te p ins tr u c tio ns .

MCL Ca rousels

T h e F C 50 0 flo w Cy to m e te r s ta r te r k it h a s :

r T w o c a r o u s e ls , e a c h w ith 32 tu b e p o s itio ns .

r A s h e e t o f b a r- c o de la b e ls , nu m b e r e d 0 1 to 99, fo r y o u to u s e to ide ntify th e c a r o u s e ls .

Ba r-Cod e La b els

Y o u c a n p u t a b a r-c o de la b e l o n e a c h s a m p le tu b e . S e e th e B A R -CO D E S P E CIF ICA T IO N S a p p e ndix .

Putting a Ba r-Cod e La b el on a Sa m p le Tub e

IMPORTANT S amp le mis id entification can occur from th e us e of incorrect, p oor q uality , d amag ed , d irty or imp rop erly p laced b ar-cod e lab els . Follow th e BAR -CO D E S PE CIFICAT IO NS to create y our b ar-cod e lab els to p revent incorrect s amp le id entification. R is k of erroneous res ults if th e b ar-cod e lab el is p laced incorrectly on s amp le tub es . T o p revent mis id entified s amp les , affix th e b ar-cod e lab el as s h ow n b elow s o th e M CL can read th e lab el.

1 Ca re fu lly a lig n th e la b e l w ith th e tu b e .

2 P r e s s th e la b e l do w n s e c u r e ly , inc lu ding

e dg e s a nd c o rne r s , w ith o u t w r ink le s o r fo lds .

b 25.4 m m ( 1.0 in.) m inim u m

c 7 .5 de g r e e s .

b

c

c

PN 624923B1 -1 4

SY STEM OVERVIEWL EA R NING TH E B A SIC OP ER A TING TECH NIQ U ES

Putting Sa m p le Tub es in a Ca rousel

Putting a Ca rousel in th e MCL

r T h e orientation of a tub e w ith a b ar-cod e lab el b d oes not matter. T h e M CL rotates th e tub e to find th e b ar-cod e lab el.

r D o not s kip tub e p os itions w ith in a p anel. T h e FC 500 flow cy tometer d oes not s kip a p rotocol in a p anel w h en a carous el tub e p os ition is emp ty . If y ou los e a s amp le, d elete th at p rotocol from th e p anel us ing th e Acq uis ition M anag er.

r Y ou can s kip a s ing le-tub e p os ition to s ep arate tw o p anels b as ed on y our tub e location s etup in Acq uis ition M anag er.

b

1 O p e n th e MCL c o v e r.

PN 624923B 1 -1 5

SY STEM OVERVIEWL EA R NING TH E B A SIC OP ER A TING TECH NIQ U ES 1

2 P ic k u p th e c a r o u s e l. Line u p th e

c a r o u s e l w ith its tu rnta b le , a nd th e n p u s h do w n. T h e c a r o u s e l is in h o m e p o s itio n w h e n th e h a ndle p o ints to w a rd th e b a c k .

3 Clo s e th e MCL c o v e r.

PN 624923B1 -1 6

SY STEM OVERVIEWL EA R NING TH E B A SIC OP ER A TING TECH NIQ U ES

Rem ov ing a Ca rousel from th e MCL

1 O p e n th e MCL c o v e r.

2 R e m o v e th e c a r o u s e l.

PN 624923B 1 -1 7

SY STEM OVERVIEWCX P SOFTW A R E SH OR TCU TS 1

1.5 CX P SOFTWARE SHORTCUTSCX P s o ftw a r e u s e s s e v e r a l s ta nda rd W indo w s te c h niq u e s fo r p r o v iding s h o r tc u ts to fu nc tio ns .

K e y b o a rd S h o r tc u tsD r a g A nd D r o pT o o lb a r B u tto ns

Key b oa rd Sh ortcuts

W h e r e Ý+X m e a ns h o ld Ý do w n a nd p r e s s X.


Ý+C Co p y

Ý+V P a s te

Ý+X Cu t

Ý+Z U ndo

Ý+Y R e do

Ý+W W o r k s p a c e P r e fe r e nc e s

Ý+T T ile S p e c ia l

Ý+P P r int

Ý+N N e w P r o to c o l

Ý+S S a v e P r o to c o l

Ý+O O p e n Lis tm o de F ile

Ý+F F o r m a t P lo t

PN 624923B1 -1 8

SY STEM OVERVIEWCX P SOFTW A R E SH OR TCU TS

Dra g And Drop

D r a g g ing a nd dr o p p ing is a s h o r tc u t m e th o d o f o p e ning , m o v ing a nd de le ting file s o r o th e r o b je c ts .

W ith in CX P s o ftw a r e th e D r a g a nd D r o p te c h niq u e c a n b e u s e d fo r s e v e r a l fu nc tio ns , w ith

s o m e o p e r a tio ns Ý c a n b e h e ld do w n to m o dify th e de fa u lt b e h a v io r.

Ý+D D u p lic a te P lo t

Ý+Q Lis tm o de Q u ic k CO MP

Ý+L V ie w Cy to m e te r Lo g

Ý+1 Cre a te ne w D o t P lo t

Ý+2 Cre a te ne w His to g r a m P lo t

Ý+3 Cre a te ne w Co nto u r P lo t

Ý+4 Cre a te ne w D e ns ity P lo t

Ý+5 Cre a te ne w O v e r la y P lo t ( CX P A na ly s is S o ftw a r e o nly )

Ý+6 Cre a te ne w T o m o g r a m P lo t ( CX P A na ly s is S o ftw a r e o nly )

Ý+7 Cre a te ne w S u r fa c e P lo t ( CX P A na ly s is S o ftw a r e o nly )

Ý+8 Cre a te ne w P r is m P lo t

Ý+9 Cre a te ne w Le g e nd P lo t

Ý+0 Cre a te ne w Info P lo t

Þ+Ì E x it CX P s o ftw a r e

É Co nte x t S e ns itiv e He lp

Í R e fr e s h S c r e e n

Ï P u b lis h to E x c e l ( o r te x t file )

Ñ S ta r t A c q u is itio n

Ò S to p A c q u is itio n

Ó P a u s e A c q u is itio n

Ô A b o r t A c q u is itio n

Dra g From … Action

Resource Ex p lorer

W orklis t D rag a s tored W orklis t to Acq uis ition M anag er.

Panels Ad d a p anel to th e current W orklis t b y d rag g ing a p anel to th e current W orklis t.

Protocols D rag a p rotocol to th e W orks p ace or Acq uis ition M anag er.

L is tmod e files D rag a file to a s eries of p lots , or w ith Ý p res s ed O NL Y th e current p lot.

H is tog ram files D rag h is tog rams to an overlay p lot.

Protocol Ex p lorer

PN 624923B 1 -1 9

SY STEM OVERVIEWCX P SOFTW A R E SH OR TCU TS 1

Toolb a r Buttons

T o o lb a r s fo r m a n e s s e ntia l p a r t o f th e CX P s o ftw a r e . T h e a p p lic a tio n e nte r e d a nd th e m o de y o u a r e o p e r a ting a t th e tim e de te r m ine s w h e th e r a p a r tic u la r T o o lb a r dis p la y is a v a ila b le .

T o o lb a r s c a n b e m o v e d u s ing th e D r a g a nd D r o p m e th o d w ith th e m o u s e a r o u nd th e CX P s o ftw a r e de s k to p o r c u s to m iz e d to s u it th e p r e fe r e nc e s o f e a c h u s e r. S e e , T o o lb a r s - Cu s to m iz e T o o lb a r s in th e R e fe r e nc e m a nu a l.

T h e T o o lb a r e le m e nts a r e :

F ILE O P T IO N S T O O LB A R

P LO T O P T I O N S T O O LB A R

R E G I O N S O P T IO N S T O O LB A R

G A T E , CO LO R , S T A T S A N D HE LP T O O LB A R

F LO W P A G E T O O LB A R

A CQ U IS I T IO N MA N A G E R T O O LB A R

A U T O MA T O R T O O LB A R

O V E R LA Y T O O LB A R ( A na ly s is o nly )

CY T O ME T E R T O O LB A R .

G ates D rag a g ate to g ate AL L p lots on a reg ion, or w ith Ý p res s ed O NL Y th e current p lot.

AutoMATOR Setup

R earrang e files R eord er files for p lay b ack us ing th e AutoM AT O R ap p lication.

Mod ify Color Preced ence

R eord er G ates D rag g ates into th e req uired p reced ence ord er.

Plots

R eg ions D rag cop ies of reg ions from one p lot to anoth er, w ith Ý p res s ed g ate log ic is als o cop ied .

Plot Imag es D rag th e current p lot imag e to a th ird p arty ap p lication (s uch as , M S Paint, Pow er

Point®).

O verlay Plot R eord er h is tog rams in th e overlay .

S tatis tics R eord er th e s tatis tics und er th e p lots .

Tile Sp ecia l

Plots R eord er p lots on th e d es ktop .

Cy tom eter Control (Cy tom eter Only )

Parameters R eord er p arameters in th e “ s elected Parameter” lis t.

Acq uisition Ma na g er

W orklis t Columns R eord er W orklis t columns to any d es ired ord er.

Dra g From … Action

PN 624923B1 -20

SY STEM OVERVIEWCR EA TING P R OTOCOL S

1.6 CREATING PROTOCOLS

1 File tt New tt New Protocol.

2 a nd Pa ra m eters b u tto n o n th e Cy to m e te r Co ntr o l A c q u is itio n S e tu p T a b .

3 Ch o o s e th e P a r a m e te r s to b e a c q u ir e d a nd .

4 If th e p a r a m e te r s h a v e c h a ng e d,

a . .

b . E nte r th e p a r a m e te r na m e a nd .

c . o n th e Cy to m e te r Co ntr o l s c r e e n.

5 Cre a te th e P lo ts r e q u ir e d fo r a c q u is itio n.

6 If a na ly s is is to b e p e r fo r m e d du r ing a c q u is itio n, c r e a te th e r e q u ir e d R e g io ns a nd G a te s ,

a nd a ls o s e le c t th e r e q u ir e d s ta tis tic s fr o m th e Ana ly sis tt Select Results o p tio n.

PN 624923B 1 -21

SY STEM OVERVIEWCR EA TING P R OTOCOL S 1

7 File tt Sa v e Protocol As to s a v e th e p r o to c o l.

8 E nte r th e s a m p le info r m a tio n into th e c o r r e c t p o s itio n in th e A c q u is itio n Ma na g e r

W o r k lis t. S e e A c q u is itio n Ma na g e r in th e R e fe r e nc e m a nu a l to m o dify th e A c q u is is tio n Ma na g e r dis p la y .

9 S e t th e lis tm o de file na m e o p tio ns fr o m th e W o r k s p a c e P r e fe r e nc e s - LMD F ile N a m e ta b .

10 S e t th e a c q u is itio n o p tio ns fr o m th e W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns ta b .

11 S e t u p th e W o r k lis t a nd r u n s a m p le s .

12 S e tu p Mo de o n th e Cy to m e te r Co ntr o l A c q u is itio n S e tu p T a b a nd a dju s t th e

ins tr u m e nt s e tting s o n th e Cy to m e te r Co ntr o l D e te c to r T a b .

13 W h e n s a tis fie d w ith th e da ta , , de s e le c t S e tu p Mo de a nd c o ntinu e a c q u is itio n.

14 O nc e s u ffic ie nt da ta is c o lle c te d, File tt Sa v e Protocol to u p da te th e p r o to c o l w ith th e

ne w ins tr u m e nt s e tting s .

PN 624923B1 -22

SY STEM OVERVIEWCR EA TING R EG IONS

1.7 CREATING REGIONSCre a te P o ly g o na l R e g io nsCre a te R e c ta ng u la r R e g io nsCre a te Q u a dr a nt R e g io nsCre a te Line a r R e g io ns

Crea te Poly g ona l Reg ions

S e e a ls o : Inte r a c tiv e P o ly g o na l R e g io n E diting in th e R e fe r e nc e m a nu a l.

1 Hig h lig h t a du a l p a r a m e te r p lo t ( c lic k o n th e title b a r ) .

2 .

3 A P o ly g o na l R e g io n c u r s o r is dis p la y e d o n s c r e e n.

4 P la c e th e c u r s o r a t th e p o int w h e r e y o u w a nt th e R e g io n to b e g in.

5 Clic k a nd r e le a s e th e m o u s e b u tto n, to fix th e p o int ( a s ta r t b o x is dis p la y e d o n s c r e e n) .

6 Co ntinu e in th is m a nne r u ntil th e de s ir e d nu m b e r o f p o ints h a s b e e n c h o s e n.

W h e n dr a w ing a R e g io n, if y o u de c ide th a t it is no t th e o ne y o u w a nt, p r e s s È b e fo r e th e r e g io n is c o m p le te d. T h e R e g io n is de le te d.

T o c h a ng e th e R e g io n ty p e , a nd s ta y in th e c u r r e nt w indo w , c h o o s e fr o m th e R e g io n m e nu o r c lic k a R e g io n s h o r tc u t b u tto n fo r w h a te v e r D u a l P a r a m e te r R e g io n ty p e y o u w is h .

PN 624923B 1 -23

SY STEM OVERVIEWCR EA TING R EG IONS 1

Crea te Recta ng ula r Reg ions

S e e a ls o : Inte r a c tiv e R e c ta ng u la r R e g io n E diting in th e R e fe r e nc e m a nu a l.

7 Clo s e th e R e g io n b y r e tu rning th e

c u r s o r to th e s ta r ting b o x a nd c lic k ing to c o nc lu de R e g io n c r e a tio n.

8 T h e R e g io n N a m e a p p e a r s o n th e p lo t.

9 S e e D o t P lo t D a ta S o u rc e to dis p la y p e r c e nta g e s w ith th e R e g io n na m e .

1 Hig h lig h t a du a l p a r a m e te r p lo t.

2 .

3 A R e c ta ng u la r R e g io n c u r s o r a p p e a r s in th e h ig h lig h te d p lo t.

4 P o s itio n th e c u r s o r to th e p o int w h e r e y o u w a nt th e R e g io n to b e g in.

PN 624923B1 -24


Crea te Qua d ra nt Reg ions

5 Clic k a nd r e le a s e th e m o u s e b u tto n. A R e c ta ng u la r R e g io n s ta r t b o x is dis p la y e d o n

s c r e e n. B y m o v ing th e m o u s e y o u c a n inc r e a s e a nd de c r e a s e th e s iz e o f th e R e c ta ng u la r R e g io n. W h e n th e de s ir e d R e c ta ng u la r R e g io n a p p e a r s , c lic k th e m o u s e b u tto n to c o m p le te th e R e g io n.

6 T h e R e g io n N a m e a p p e a r s o n th e p lo t.

1 Hig h lig h t a du a l p a r a m e te r p lo t.

2 .

3 A c r o s s h a ir c u r s o r is dis p la y e d in th e c u r r e nt w indo w .

4 P o s itio n th e c u r s o r o n th e p lo t w h e r e y o u w a nt th e inte r s e c tio n p o int o f th e r e g io n a nd

to s h o w th e q u a dr a nt line s ( c r o s s h a ir ) .

5 Mo v e th e r e g io ns to th e r e q u ir e d p o s itio n, if ne e de d, th e n to s e t th is p o s itio n.

PN 624923B 1 -25


W h e n dr a w ing a R e g io n, if y o u de c ide th a t it is no t th e o ne y o u w a nt, p r e s s È b e fo r e th e R e g io n is c o m p le te d. T h e R e g io n is de le te d.

N ote: Q u a dr a nt R e g io n is a s p e c ia l c a s e o f r e c ta ng u la r R e g io ns - fo u r R e c ta ng u la r R e g io ns a r e s im u lta ne o u s ly c r e a te d w ith o ne m o u s e c lic k a nd e x is t th e r e a fte r a s fo u r inde p e nde nt R e c ta ng u la r R e g io ns ( w ith th e e x c e p tio n th a t th e y do no t s a tis fy A u to m a tic G a te Cre a tio n) . D e le ting o ne o r m o r e s till le a v e s r e m a ining R e g io ns inta c t if u s ing R e g io n/V ie w /Mo dify D e le te . I f y o u u s e th e g r a p h ic a l q u a dr a nt de le te , th e n a ll fo u r Q u a dr a nt R e g io ns a r e de le te d a s a g r o u p .

Rep ositioning a Qua d ra nt Reg ion

S e e a ls o : Inte r a c tiv e Q u a dr a nt R e g io n E diting in th e R e fe r e nc e m a nu a l.

6 CX P s o ftw a r e s h o w s th e Q u a dr a nt's na m e in e a c h o f its fo u r R e g io ns .

1 Mo u s e o v e r th e Q u a dr a nt r e g io n line s to g e t th e , o r c u r s o r.

2 R e p o s itio n th e r e g io n.

a . + h o ld a nd dr a g to m o v e th e v e r tic a l div ide r.

b . + h o ld a nd dr a g to m o v e th e h o r iz o nta l div ide r.

c . + h o ld a nd dr a g to m o v e th e Q u a dr a nt r e g io n.

3 R e le a s e th e m o u s e b u tto n.

PN 624923B1 -26


Crea te Linea r Reg ions

T h is o p tio n r e q u ir e s tw o p o ints to b e s e t g iv ing a n u p p e r a nd lo w e r c h a nne l b o u nda r y fo r e v e r y R e g io n.

S e e a ls o ( in th e R e fe r e nc e m a nu a l) :Inte r a c tiv e Line a r R e g io n E ditingS ta tu s B a r ( Line a r )X Co o rdina teInte g r a l

1 Hig h lig h t a s ing le p a r a m e te r h is to g r a m .

2 .

A Line a r R e g io n c u r s o r a p p e a r s in th e c u r r e nt w indo w .

3 P o s itio n th e c u r s o r o n th e c h o s e n p lo t to th e p o int w h e r e y o u w a nt th e R e g io n to b e g in.

Clic k , tw o v e r tic a l line s w ith a h o r iz o nta l b a r a p p e a r s .

4 Mo v e th e c u r s o r le ft o r r ig h t, u p o r do w n u ntil th e de s ir e d e nd p o int o f th e R e g io n is

r e a c h e d. to a c c e p t th e s e tting s a nd a nc h o r th e r e g io n.

W h e n dr a w ing a R e g io n, if y o u de c ide th a t it is no t th e o ne y o u w a nt, p r e s s È b e fo r e th e R e g io n is c o m p le te d. T h e R e g io n is de le te d.

A fte r s e tting th e R e g io n p o s itio n, th e lo ng v e r tic a l line s a r e r e p la c e d b y s h o r t c h a nne l m a r k e r s to p r e v e nt c lu tte r ing o f th e p lo t dis p la y .

T h e h o r iz o nta l b a r is p o s itio ne d a p p r o x im a te ly a t th e h o r iz o nta l c u r s o r p o s itio n w h e n th e r e g io n is s e t.

PN 624923B 1 -27


Crea te Multip le Linea r Reg ions

S e e a ls o : Inte r a c tiv e Mu lti-Line a r R e g io n E diting in th e R e fe r e nc e m a nu a l.

1 Hig h lig h t a s ing le p a r a m e te r h is to g r a m a nd .

2 A P a r a lle l c u r s o r a p p e a r s in th e c u r r e nt w indo w .

3 P o s itio n th e c u r s o r o n th e c h o s e n p lo t to th e p o int w h e r e y o u w a nt th e R e g io n to b e g in.

Clic k , tw o v e r tic a l line s w ith a h o r iz o nta l b a r is dis p la y e d. A s to p b o x a p p e a r s in th e r ig h t h a nd to p c o rne r o r th e w indo w .

4 Mo v e th e c u r s o r le ft o r r ig h t, u p o r do w n u ntil th e de s ir e d e nd p o int o f th e R e g io n is

r e a c h e d. to a c c e p t th e s e tting s a nd a nc h o r th e r e g io n.

5 R e p e a t th is p r o c e du r e u ntil a ll R e g io ns a r e s e t.

6 W h e n y o u h a v e dr a w n a ll o f y o u r Mu lti-Line a r R e g io ns , p r e s s È o r m o v e th e c u r s o r

o nto th e s to p b o x in th e r ig h t h a nd to p c o rne r, c lic k . T h is e nds th e Mu lti-Line a r s e q u e nc e .

PN 624923B1 -28

SY STEM OVERVIEWCR EA TING G A TES

1.8 CREATING GATES

Crea te AutoGa te

N ote: If a s to p c o u nt is u s e d o n a p lo t th a t c o nta ins a n A u to G a te r e g io n, th e s to p c o u nt is no t e x a c t.

1 in th e p lo t y o u w a nt to c r e a te a n

A u to G a te o n.

2 o r .

3 o n th e p o p u la tio n to A u to G a te .

PN 624923B 1 -29

SY STEM OVERVIEWCR EA TING G A TES 1

4 T o s p e c ify th e A u to G a te S e ns itiv ity a nd

T r a v e l,

a . th e r e g io n.

b . Clic k th e r ig h t m o u s e b u tto n o n

th e r e g io n a nd Reg ion

Prop erties.

c . S p e c ify th e A u to G a ting S e ns itiv ity a nd Co nto u r T r a v e l, if ne c e s s a r y ,

a nd .

5 R ig h t m o u s e c lic k o n th e p lo t y o u w a nt

to a s s ig n th e A u to G a te to a nd s e le c t F o r m a t to dis p la y th e P lo t P r o p e r tie s dia lo g .

6 th e g a ting dr o p do w n lis t a nd

s e le c t th e A u to G a te to g a te th e p lo t o n th e A u to G a te .

7 R e p e a t s te p s 5 a nd 6 to a s s ig n th e A u to G a te to o th e r p lo ts o r Ap p ly g a te to a ll p lots.

N ote: I f y o u s e le c t Ap p ly g a te to a ll p lots y o u m u s t g o b a c k to th e p lo t th a t c o nta ins th e A u to G a te a nd u na s s ig n th e A u to G a te fr o m th e p lo t.

PN 624923B1 -30

SY STEM OVERVIEWCR EA TING G A TES

Conv ert a Poly g ona l Reg ion to a n AutoGa te Reg ion

Autom a tic Ga te Crea tion

1 r e g io n to m a k e it a c tiv e .

2 Clic k th e r ig h t m o u s e b u tto n o n th e r e g io n a nd Reg ion Prop erties.

3 e ith e r Ellip tica l o r Contour in th e P o ly g o na l R e g io n P r o p e r tie s s c r e e n.

T h is o p tio n a llo w s y o u to c r e a te a ne w g a te a u to m a tic a lly w h e n a ne w R e g io n is dr a w n. If th is o p tio n is no t s e le c te d y o u m u s t u s e Ana ly sis tt Crea te Mod ify Ga tes to c r e a te g a ting lo g ic .

R e g io ns c o p ie d u s ing Ý+D r a g

a nd D r o p into a p lo t a s s ig n th e r e g io n a s a g a te .

PN 624923B 1 -31

SY STEM OVERVIEWCR EA TING F L OW P A G ES 1

1.9 CREATING FLOWPAGES

Flow PAGE Ex a m p le

B e lo w is a ty p ic a l e x a m p le o f a F lo w P A G E . T h is is a s c r e e n v ie w o nly . A fu ll p r inte d p a g e o u tp u ts a t a m u c h h ig h e r r e s o lu tio n. S e e a ls o : F lo w P A G E Me nu in th e U s ing CX P S o ftw a r e c h a p te r in th e R e fe r e nc e m a nu a l.

1 to c r e a te a b la nk F lo w P A G E .

2 to ins e r t a p ic tu r e .

3 Ins e r t F lo w p a g e P lo ts .

4 Ins e r t a F lo w P A G E S ta tis tic s T a b le .

5 Ins e r t a F lo w P A G E T e x tb o x .

6 Clic k th e r ig h t m o u s e b u tto n o n a ny

o b je c t o n th e F lo w P A G E to s e le c t a dditio na l fo r m a tting o p tio ns .

PN 624923B1 -32

SY STEM OVERVIEWCR EA TING P A NEL S

Delete Flow PAGES

1.10 CREATING PANELST h e P a ne l W iz a rd a llo w s y o u to c r e a te a P a ne l fo r u s e in th e A c q u is itio n Ma na g e r.

1 in th e u p p e r le ft c o rne r o f th e

F lo w P A G E w indo w to b e de le te d a nd

Close.

N ote: F lo w P A G E S in a lo c k e d p r o to c o l c a n no t b e ins e r te d, de le te d o r e dite d.

1 File tt New Pa nel to s ta r t th e P a ne l

W iz a rd.

2 E nte r th e N e w P a ne l na m e .

3 S e le c t th e N u m b e r o f T e s ts r e q u ir e d.

T h e de fa u lt v a lu e is 1 a nd th e m a x im u m is 32.

4 Ex p ort th e results of th is p a nel to th e Rep ort Genera tor if y o u w a nt a P a tie nt P a ne l

R e p o r t g e ne r a te d w h e n y o u r u n th is p a ne l. S e e P a ne l R e p o r t in th e R e fe r e nc e m a nu a l.

PN 624923B 1 -33

SY STEM OVERVIEWCR EA TING P A NEL S 1

5 .

6 Ch o o s e y o u r P r o to c o l S e tting s .

U s e P lo ts a nd G a te s fr o m P r e v io u s T e s tU s e P lo ts a nd G a te s fr o m P r o to c o lU s e Ins tr u m e nt S e tting s

7 Ch o o s e y o u r R e g io n S e tting s .

U s e R e g io ns fr o m P r o to c o lCa r r y R e g io ns fr o m P r e v io u s P r o to c o lU s e R e g io ns fr o m T h is P r o to c o l:

.

8 Ch o o s e y o u r Ins tr u m e nt S e tting s .

U s e Ins tr u m e nt S e tting s fr o m P r e v io u s T e s tU s e Ins tr u m e nt S e tting s fr o m E x te rna l F ile

9 D o u b le o n p a r a m e te r na m e s u nde r Prob e/Sta in na m e to a s s ig n th e P a r a m e te r N a m e s .

10 R e p e a t s te p s 6 th r o u g h 9 fo r e a c h te s t.

PN 624923B1 -34

SY STEM OVERVIEWCR EA TING P A NEL S

Use Plots a nd Ga tes from Prev ious Test

A llo w s y o u to u s e G a te s a nd P lo ts fr o m a p r e v io u s T e s t b u t is no t a c tiv e fo r th e fir s t T e s t w ith in a P a ne l.

T h is o p tio n is no t a v a ila b le if Use p lots a nd g a tes from p rotocol is c h o s e n.

Use Plots a nd Ga tes from Protocol

A llo w s y o u to s e le c t P lo ts a nd G a te s fr o m a s p e c ific P r o to c o l, c h o s e n fr o m th e dr o p do w n lis t b o x .

T h is o p tio n is no t a v a ila b le if Use p lots a nd g a tes from p rev ious test is c h o s e n.

Use Instrum ent Setting s

U s e s th e ins tr u m e nt s e tting s fr o m th e s e le c te d P r o to c o l. I f th e c h e c k b o x is dis a b le d, th is a llo w s th e Instrum ent Setting s b e lo w to b e a c tiv e .

Use Reg ions from Protocol

T h is o p tio n lo a ds a ll r e g io ns s to r e d w ith in th e c u r r e nt p r o to c o l.

Ca rry Reg ions from Prev ious Protocol

S e le c ting th is o p tio n a llo w s r e g io n p o s itio ns to b e c a r r ie d fr o m th e p r e v io u s te s t in a p a ne l. A ny r e g io ns o f th e s a m e ty p e a nd dr a w n o n th e s a m e p a r a m e te r s a r e c a r r ie d.

Use Reg ions from Th is Protocol:

T h is o p tio n lo a ds th e r e g io ns a nd r e g io n p o s itio ns fr o m th e s e le c te d p r o to c o l.

Use Instrum ent Setting s from Prev ious Test

U s e s th e Ins tr u m e nt s e tting s fr o m th e p r e v io u s T e s t b u t is no t a c tiv e fo r th e fir s t T e s t w ith in a P a ne l.

T h is o p tio n is no t a v a ila b le if Use instrum ent setting s from ex terna l file is s e le c te d.

Use Instrum ent Setting s from Ex terna l File

U s e Ins tr u m e nt S e tting s fr o m a n E x te rna l F ile s u c h a s Lis tm o de o r P r o to c o l [ S e tting s ] file s th a t y o u c h o o s e fr o m th e dr o p do w n lis t b o x .

T h is o p tio n is no t a v a ila b le if Use instrum ent setting s from p rev ious test is s e le c te d.

Pa ra m eter Na m es

A llo w s y o u to c h o o s e th e P a r a m e te r N a m e s fo r e a c h tu b e w ith in a P a ne l.

11 .

PN 624923B 1 -35

SY STEM OVERVIEWCR EA TING W OR K L ISTS 1

1.11 CREATING WORKLISTS

N ote: I f no W o r k lis t is v is ib le , View tt Acq uisition Ma na g er to dis p la y th e W o r k lis t p a ne .

N ote: U nle s s th e c u r r e nt W o r k lis t h a s b e e n s a v e d u s ing th e Sa v e Work list b u tto n w ith in th e A CQ U IS I T IO N MA N A G E R T O O LB A R , c u r r e nt w o r k lis t s e tting s a r e no t a v a ila b le .

Work list Pa nel

1 to c r e a te a ne w W o r k lis t o r to c le a r c u r r e nt W o r k lis t fr o m th e A c q u is itio n

Ma na g e r s c r e e n.

2 D r a g A nd D r o p p a ne ls a nd p r o to c o ls fr o m th e R e s o u rc e E x p lo r e r to th e A c q u is itio n

Ma na g e r . Info r m a tio n a dde d in th is w a y a lw a y s a p p e a r s a t th e e nd o f th e W o r k lis t.

3 A fte r a ll th e info r m a tio n h a s b e e n a dde d, y o u c a n u s e D r a g A nd D r o p to c h a ng e th e

o rde r.

1 .

2 S e le c t w h ic h p a ne l y o u w is h to o p e n

a nd .

PN 624923B1 -36

SY STEM OVERVIEWCR EA TING W OR K L ISTS

Work list Test

to ins e r t a s ing le R o w a t th e e nd o f th e c u r r e nt P a ne l.

3 A fte r a ll th e info r m a tio n h a s b e e n a dde d, y o u c a n u s e D r a g A nd D r o p to c h a ng e th e

o rde r.

Y o u c a n u s e th is o p tio n to a dd a no th e r P a ne l to a n e x is ting W o r k lis t.

PN 624923B 1 -37

SY STEM OVERVIEWU SING TH E ONL INE H EL P SY STEM 1

1.12 USING THE ONLINE HELP SY STEMY o u r Cy to m ic s F C 50 0 s y s te m p r o v ide s a n o nline He lp s y s te m th a t a llo w s y o u to s e a rc h fo r info r m a tio n o n s p e c ific s y s te m - r e la te d to p ic s th r o u g h th e Contents, Ind ex , and Sea rch o p tio ns .

T h e He lp s y s te m is a n e le c tr o nic v e r s io n o f th e Ins tr u c tio ns F o r U s e Ma nu a l, R e fe r e nc e Ma nu a l, a nd S p e c ia l P r o c e du r e s Ma nu a l. It inc lu de s a ta b le o f c o nte nts , a n inde x fo r finding info r m a tio n q u ic k ly , a nd a g lo s s a r y o f de finitio ns .

W h e n y o u a c c e s s th e He lp m e nu , th e r e a r e th r e e o p tio ns a v a ila b le : CX P Help , Ab out..., a nd

Online Sup p ort.

Access Online Help

How to Use Help

U s e o ne o f th e s e o p tio ns to a c c e s s th e o nline He lp s y s te m .

.

Help tt CX P Help .

É.

N ote: I f y o u c a nno t a c c e s s th e o nline He lp s y s te m , c o nta c t y o u r B e c k m a n Co u lte r R e p r e s e nta tiv e .

o n th e He lp N a v ig a tio n B a r.

Info r m a tio n a b o u t th e He lp w indo w a nd ins tr u c tio ns fo r u s ing h e lp a r e dis p la y e d in th e na v ig a tio n p a ne .

b T o o lb a rc N a v ig a tio n B a rd T o p ic P a nee N a v ig a tio n P a ne

PN 624923B1 -38

SY STEM OVERVIEWP ER F OR M A NCE CH A R A CTER ISTICS

Help Menu

CX P Help

O p e ns th e F C 50 0 CX P o nline h e lp .

Ab out...

D is p la y s :

r CX P s o ftw a r e c o p y r ig h t

r CX P s o ftw a r e v e r s io n info r m a tio n

r R u ntim e P r o to c o l o r N e w P r o to c o l ( w h e n lis tm o de file is r e p la y e d) o r Lo c k e d P r o to c o ls .

r th a t ta k e s y o u to a lis t o f c u r r e nt a na ly s is a lg o r ith m s a nd th e ir de s c r ip tio ns .

If y o u a r e a s k ing fo r te c h nic a l s u p p o r t b y te le p h o ne , fa x , o r e m a il, p le a s e q u o te th e CX P s o ftw a r e b u ild nu m b e r, w h ic h is a c c e s s ib le fr o m m e nu Help tt Ab out..., a nd e ns u r e y o u k no w th e R o m lo c k nu m b e r w h ic h is w r itte n o n th e b o dy o f th e R o m lo c k .

Online Sup p ort

r Beck man Cou lter Web Site

If y o u h a v e a n Inte rne t c o nne c tio n a nd y o u c lic k th e m o u s e o n h ttp ://w w w .b e c k m a nc o u lte r.c o m y o u c a n a c c e s s th e B e c k m a n Co u lte r W e b S ite th r o u g h y o u r W e b B r o w s e r p r o g r a m .

r ACS Web Site

If y o u h a v e a n Inte rne t c o nne c tio n a nd y o u c lic k th e m o u s e o n h ttp ://w w w .a p p lie dc y to m e tr y .c o m y o u c a n a c c e s s th e A p p lie d Cy to m e tr y S y s te m s W e b S ite th r o u g h y o u r W e b B r o w s e r p r o g r a m .

Y o u c a n th e n a c c e s s th e la te s t info r m a tio n c o nc e rning th e u p -to -da te s o ftw a r e r e le a s e s a nd o th e r ite m s o f inte r e s t.

1.13 PERFORMANCE CHARACTERISTICSR e fe r to th e p a c k a g e ins e r t o f th e Q u a lity Co ntr o l Ma te r ia ls lis te d in th e R e fe r e nc e m a nu a l fo r th e p e r fo r m a nc e c h a r a c te r is tic s fo r th e p r e p a r a tio n m e th o d y o u a r e u s ing .

PN 624923B 2-1

22DAILY ROUTINE

2.1 BEFORE Y OU BEGINT h is c h a p te r e x p la ins th e da ily s ta r tu p p r o c e du r e s . B e fo r e do ing th e s e p r o c e du r e s :

2.2 DAILY STARTUPP e r fo r m th e fo llo w ing s te p s to s ta r t u p th e s y s te m . I f y o u h a v e s e t u p CY T O ME T E R A U T O S T A R T U P a nd th e Cy to m e te r is r u nning , s k ip a h e a d to Ch e c k th e P o w e r S u p p ly .

1 R e a d th e O P E R A T I O N P R IN CIP LE S c h a p te r. U s ing y o u r s y s te m is e a s ie r if y o u h a v e a

g e ne r a l u nde r s ta nding o f h o w it w o r k s .

2 R e a d th e S Y S T E M O V E R V I E W c h a p te r. It c o nta ins ins tr u c tio ns fo r

r CY T O ME T E R CO N T R O LS A N D IN D ICA T O R S

r LE A R N I N G T HE B A S IC O P E R A T IN G T E CHN IQ U E S .

3 R e a d e a c h p r o c e du r e e ntir e ly .

4 If c o nditio ns c a u s e s ta tic c h a r g e to e x is t in y o u r la b , b e s u r e to p r o p e r ly g r o u nd y o u r s e lf

b e fo r e to u c h ing th e ins tr u m e nt.

5 S h u tdo w n a nd r e s ta r t th e s y s te m c o m p u te r o nc e p e r da y to a llo w th e v ir u s p r o te c tio n

p r o g r a m to r u n. S e e P o w e r th e Co m p u te r a nd Cy to m e te r O F F a nd P o w e r th e Co m p u te r a nd Cy to m e te r O N . D o no t s ta r t a fu ll dis k v ir u s s c a n w h ile r u nning CX P s o ftw a r e .

PN 624923B2-2

DAILY ROUTINED A IL Y STA R TU P

Ch eck Wa ste a nd Rea g ent Lev els

1 E m p ty th e w a s te c o nta ine r a nd v e r ify

tu b ing is c o nne c te d to th e c a p .

2 Ch e c k th e s h e a th flu id le v e l a nd fill th e

s h e a th c o nta ine r if ne c e s s a r y .

PN 624923B 2-3

DAILY ROUTINED A IL Y STA R TU P 2

Pow er th e Com p uter a nd Cy tom eter ON

Turning On Pow er

3 Ch e c k th e c le a ning a g e nt fill le v e l b

a nd fill th e c le a ning a g e nt c o nta ine r if ne c e s s a r y .

b

T u rn o n th e s y s te m a t th e c o m p u te r.

PN 624923B2-4


Log g ing Onto Wind ow s Softw a re

Log g ing Onto CX P Softw a re

W h e n th e Log on to Wind ow s s c r e e n a p p e a r s ,

a . Ch e c k th a t th e User na m e fie ld dis p la y s Administrator o r y o u r W indo w s lo g - in u s e r na m e . If no t, ty p e in y o u r u s e r na m e .

N ote: W h e n fir s t ins ta lling th e s o ftw a r e , th e User na m e fie ld m u s t dis p la y Administrator.

b . E ns u r e th e c h e c k b o x is u nc h e c k e d.

c . E nte r y o u r P a s s w o rd a nd

.

N ote: I f y o u r c o m p u te r is p a r t o f a ne tw o r k , y o u m a y ne e d to e nte r th e U s e r na m e a nd P a s s w o rd a s s ig ne d b y y o u r ne tw o r k m a na g e r.

1 to s ta r t th e

s o ftw a r e a nd p o w e r u p th e Cy to m e te r.

A llo w a b o u t 40 m inu te s to w a r m u p th e s y s te m b e fo r e p e r fo r m ing Q C o r r u nning s a m p le s .

PN 624923B 2-5


2 A t th e CX P Cy tom eter Sta rtup Wiz a rd [Pa g e

1 of 2] s c r e e n:

a . Hig h lig h t y o u r U s e r I D .

b . E nte r y o u r P a s s w o rd.

c . .

N ote: F o r a dditio na l info r m a tio n a b o u t th is s c r e e n, s e e MU LT I-U S E R S IG N O N . I f y o u ne e d to s e t u p U s e r I D s , s e e U s e r A dm inis tr a tio n.

3 S e le c t a p r o to c o l, if ne e de d.

4 .

5 D u r ing s y s te m s ta r tu p , th e fo llo w ing s e r ie s o f Cy to m e te r s ta tu s m e s s a g e s a r e dis p la y e d.

T h e s ta r tu p c y c le inc lu de s a p r im e c y c le .

S tartup in P rocessV erificationInitializ ationA waiting S ample

PN 624923B2-6


Ch eck th e Pow er Sup p ly

1 O p e n th e P o w e r S u p p ly do o r a nd c h e c k

th e W A T E R T R A P , A IR F ILT E R , a nd V A Cu u m F ILT E R .

Ca ll y o u r B e c k m a n Co u lte r R e p r e s e nta tiv e if:

r T h e T R A P is > 1/3 fu ll.

r T h e F ILT E R S h a v e a ny flu id.

2 Ch e c k th e S Y S te m V A Cu u m g a u g e . I f it

r e a ds ≥−17 in. Hg , c a ll y o u r B e c k m a n Co u lte r R e p r e s e nta tiv e .

3 Ch e c k th e S Y S te m P R E S S u r e g a u g e . I f

it do e s no t r e a d b e tw e e n 28 a nd 32 p s i, do th e fo llo w ing :

WATER TRAP

VAC TRAP

AIR FILTER

VAC

FILTER

SY S VAC

SY S PRESS

30 PSI

PRESSADJ

-1 0

-5

0

-1 5

-2 0

-2 5

-3 0

> -1 7 " H g

30 ± 2 PS I

5 0

0

2 0

1 0

4 0

6 0

3 0

PN 624923B 2-7


a . P u ll th e P R E S S u r e A D J u s te r k no b o u t to w a rd y o u .

b . A dju s t th e p r e s s u r e to 30 ± 2 p s i.

b T o de c r e a s e , tu rn to th e le ft.

c T o inc r e a s e , tu rn to th e r ig h t.

c . P u s h in o n th e k no b to lo c k it into p la c e .

PU

LL TO ADJU

ST

PU

LL TO ADJU

ST

b

c

PU

LL TO ADJU

ST

PN 624923B2-8


Ad d itiona l Sta rt Up Ch eck s

4 Ch e c k th e V A Cu u m T R A P .

If it is > 1/4 fu ll o f flu id CLE A N T HE V A CU U M T R A P .

5 Clo s e th e P o w e r S u p p ly do o r.

6 R e c o rd th e s ta r tu p c h e c k s o n th e Ma inte na nc e Lo g o r o n th e A c tio n Lo g S h e e t.

1 Ch e c k th a t th e MCL v o r te x fu nc tio n m ix e s s a m p le s b y r u nning a b la nk s a m p le .

2 R e fe r to th e m a nu a ls th a t c a m e w ith y o u r P r inte r to :

r P e r fo r m P r inte r dia g no s tic s .

r Ch e c k th a t th e r e is a n a de q u a te p a p e r s u p p ly in th e P r inte r.

r Ch e c k ink c a r tr idg e s if y o u h a v e a c o lo r P r inte r a nd r e p la c e if ne c e s s a r y .

WATER TRAP

VAC TRAP

AIR FILTER

VAC

FILTER

SY S VAC

SY S PRESS

30 PSI

PRESSADJ

PN 624923B 2-9

DAILY ROUTINED A IL Y SH U TD OW N 2

2.3 DAILY SHUTDOWN

Wh en to Sh ut Dow n th e Cy tom eter

r S h u t do w n th e ins tr u m e nt a t le a s t o nc e a da y, e v e n if it is inte nde d fo r u s e 24 h o u r s p e r da y.

r Le a v e th e ins tr u m e nt s h u t do w n fo r a t le a s t 30 m inu te s b e fo r e r e s ta r ting .

r S h u tdo w n a nd r e s ta r t th e s y s te m c o m p u te r o nc e p e r da y to a llo w th e v ir u s p r o te c tio n p r o g r a m to r u n. S e e P o w e r th e Co m p u te r a nd Cy to m e te r O F F a nd P o w e r th e Co m p u te r a nd Cy to m e te r O N . D o no t s ta r t a fu ll dis k v ir u s s c a n w h ile r u nning CX P s o ftw a r e .

Before Perform ing Sh ut Dow n

Pow er th e Com p uter a nd Cy tom eter OFF

1 P e r fo r m th e R o u tine Cle a ning P r o c e du re in th e S p e c ia l P r o c e du re s Ma nu a l.

2 P e r fo r m th e V a c u u m Line Cle a ning P r o c e du re in th e S p e c ia l P r o c e du r e s Ma nu a l.

1 to p u t th e Cy to m e te r in th e Idle m o de .

2 in a ll o p e n w indo w s .

3 o r .

PN 624923B2-1 0

DAILY ROUTINED A IL Y SH U TD OW N

4 tt to tu rn o ff th e Cy to m e te r.

5 tt to

s h u tdo w n th e W o r k s ta tio n.

6 W h e n th e Sh ut Dow n Wind ow s s c r e e n

a p p e a r s :

E ns u r e Sh ut d ow n or Sta nd b y a p p e a r s b e lo w Wh a t d o y ou w a nt th e com p uter to

d o?

7 .

T h e Cy to m e te r a u to m a tic a lly tu rns o ff.

8 T u rn o ff th e m o nito r a nd P r inte r s e p a r a te ly .

PN 624923B 2-1 1

DAILY ROUTINED A IL Y SH U TD OW N 2

After Instrum ent Sh ut Dow n

1 W ip e do w n a ll e x p o s e d s u r fa c e s w ith 10 % b le a c h s o lu tio n a nd th e n 7 0 % e th a no l. P a y

s p e c ia l a tte ntio n to th e S a m p ling a r e a .

2 K e e p th e s y s te m s h u t do w n fo r 30 m inu te s . B e fo r e r u nning s a m p le s , do th e da ily s ta r tu p

a nd q u a lity c o ntr o l p r o c e du r e s .

R e m inde r :

Cle a n th e a ir filte r s o nc e a w e e k .

Cle a n th e s a m p le p r o b e a nd s a m p le h e a d o nc e a w e e k .

Cle a n th e s h e a th flu id c o nta ine r o nc e a m o nth .

Cle a n th e c le a ning a g e nt c o nta ine r e v e r y 60 da y s .

3 R e c o rd da ily s h u tdo w n a nd c le a ning o n th e Ma inte na nc e Lo g o r o n th e A c tio n Lo g

S h e e t.

PN 624923B2-1 2

DAILY ROUTINEEX TEND ED SH U TD OW N

2.4 EX TENDED SHUTDOWNIf y o u inte nd to le a v e th e ins tr u m e nt in th e s h u tdo w n s ta te fo r a n e x te nde d a m o u nt o f tim e :

1 R e m o v e th e c le a ning a g e nt a nd s h e a th

flu id c o nta ine r s .

2 R ins e th e ins ide o f b o th c o nta ine r s w ith

w a te r.

PN 624923B 2-1 3

DAILY ROUTINEEX TEND ED SH U TD OW N 2

3 R e p la c e th e c le a ning a g e nt a nd inte rna l

s h e a th flu id c o nta ine r s . F ill b o th c o nta ine r s w ith w a te r ins te a d o f r e a g e nt.

4 P e r fo r m th e R o u tine Cle a ning

P r o c e du r e . U s e ta p w a te r in tu b e s 1-4 ins te a d o f b le a c h in tu b e 1 a nd Is o F lo w s h e a th flu id in tu b e s 2-4.

5 W h e n y o u s ta r t u p th e ins tr u m e nt fo r

th e fir s t tim e a fte r th e e x te nde d s h u tdo w n,

r Cle a n th e s h e a th flu id c o nta ine r

r Cle a n th e c le a ning a g e nt c o nta ine r

r F ill th e s h e a th flu id c o nta ine r

r F ill th e c le a ning a g e nt c o nta ine r

r P e r fo r m th e R o u tine Cle a ning P r o c e du r e

r P e r fo r m D A ILY Q C b e fo r e r u nning s a m p le s .

PN 624923B2-1 4

DAILY ROUTINECY TOM ETER A U TO STA R TU P

2.5 CY TOMETER AUTO STARTUPY o u c a n s e t u p th e s y s te m to a u to m a tic a lly S ta r tu p o r S h u tdo w n th e Cy to m e te r. T h e c o m p u te r m u s t b e o n w ith W indo w s r u nning to a llo w A u to S ta r tu p to r u n.

1 U s ing W indo w s E x p lo r e r, do u b le c lic k

o n C:\CX P \T a s k s .e x e .

2 .

3 O n th e T a s k s c r e e n, e nte r C:\CX P in th e

Sta rt in: fie ld.

Y o u c a n a ls o e nte r c o m m e nts a nd s e t a p a s s w o rd, a lth o u g h th e y a r e no t r e q u ir e d.

PN 624923B 2-1 5

DAILY ROUTINECY TOM ETER A U TO STA R TU P 2

4 to s c h e du le w h e n y o u

w a nt th e S ta r tu p o r S h u tdo w n ta s k to o c c u r.

a nd e nte r y o u r s e tting s .

5 to s p e c ify a dditio na l

s e tting s .

6 If y o u w a nt to s e t u p a u to m a tic Cy to m e te r s h u tdo w n, a nd p e r fo r m

s te p s 3 th r o u g h 5.

PN 624923B2-1 6

DAILY ROUTINECY TOM ETER A U TO STA R TU P

PN 624923B 3-1

33QUALITY CONTROL

3.1 INTRODUCTIONP e r fo r m th e fo llo w ing q u a lity c o ntr o l c h e c k s to e ns u r e th a t y o u r s y s te m is w o r k ing a c c u r a te ly a nd p r e c is e ly . T h e p r o to c o ls ne e de d fo r th e s e q u a lity c o ntr o l ( Q C) p r o c e du r e s a r e inc lu de d w ith y o u r s y s te m S o ftw a r e . T h e p a c k a g e ins e r ts fo r y o u r Q u a lity Co ntr o l Ma te r ia ls h a v e ins tr u c tio ns fo r e s ta b lis h ing y o u r la b o r a to r y 's no r m a l r a ng e s fo r da ily u s e .

Y o u m u s t r e -e s ta b lis h y o u r la b o r a to r y 's r a ng e s :

r W h e n y o u u s e a ne w lo t o f flu o r o s p h e r e s .

r W h e ne v e r a m a jo r p a r t o f th e s y s te m h a s b e e n s e r v ic e d o r r e p la c e d ( fo r e x a m p le , la s e r a lig nm e nt o r r e p la c e m e nt, P MT r e p la c e m e nt) .

In a dditio n to do ing th e da ily q u a lity c o ntr o l p r o c e du r e in th is c h a p te r, y o u s h o u ld m a k e a q u a lity c o ntr o l c h e c k fo r th e s p e c ific a p p lic a tio ns y o u a r e r u nning .

3.2 QC PROCESSEST h e c h a r t b e lo w s h o w s w h ic h Q C m a te r ia ls a r e ne e de d fo r e a c h Q C p r o c e s s .

QC Process QC Ma teria l Used

V erify fluid ics and las er alig nment

Flow -Ch eck™ Fluoros p h eres (Flow -Ch eck 67 5 Fluoros p h eres , Flow -Ch eck 7 7 0 Fluoros p h eres ). V erify H PCV vers us exp ected value. E xp ort res ults to th e Q C D atab as e and review th e Q C d ata.

Ad jus t h ig h voltag e and g ain for a g iven ap p lication

Flow -S et™ Fluoros p h eres (Flow -S et 67 5 Fluoros p h eres , Flow -S et 7 7 0 Fluoros p h eres ). As certain targ et mean p os ition b as ed up on ap p lication and ad jus t h ig h voltag e and g ain d aily to th at targ et. E xp ort res ults to th e Q C D atab as e and review th e Q C d ata.

M onitor ins trument linearity

Immuno-Brite™ Fluoros p h eres .

Perform ab s olute counts Flow -Count™ Fluoros p h eres .

Ad jus t color comp ens ation for a g iven ap p lication

For AutoS etup ap p lications , Cy to-Comp ™ Cells or w h ole b lood s tained w ith Q uickCO M P™ 2 or Q uickCO M P 4 kit. U s e s ing le color s tained s amp les w ith each fluoroch rome us ed in y our ap p lication. For examp le: CD 45-FIT C, CD 45-PE , CD 45-E CD , CD 45-PC5 or CD 45-PC7 .

V erify correct s etting s w ith an ap p lication Control

U p d ate th e control p rotocol w ith th e s etting s d erived from ab ove. R un a b iolog ical control eq uivalent to th e ap p lication, s uch as Immuno-T rol™ Cells , Immuno-T rol L ow Cells , Cy to-trol™ Control Cells , S tem-T rol™ Control Cells , or a normal w h ole b lood . E xp ort res ults to th e Q C D atab as e and review th e Q C d ata.

PN 624923B3-2

QUALITY CONTROLD A IL Y Q C

3.3 DAILY QCD a ily Q C c o ns is ts o f:

r P r e p a r ing A u to S e tu p S a m p le s

r R u nning th e A u to S e tu p S c h e du le r

r R u nning th e A u to S e tu p I I W iz a rd

Prep a ring AutoSetup Sa m p les

IMPORTANT R is k of erroneous res ults if th e Cy tometer h as b een id le for an extend ed p eriod of time or y ou h ave jus t p erformed D aily S tartup . T o ens ure correct res ults , p erform a p rime after:

r D aily S tartup .

r T h e Cy tometer h as b een id le for an extend ed p eriod of time.

r Y ou p lace a new carous el on th e M CL and lig h t s catter s ig nals ap p ear ab normal.

1 P r e p a r e th e a p p r o p r ia te F lo w -Ch e c k flu o r o s p h e r e s fo r th e a p p lic a tio ns y o u ne e d to r u n.

T h e flu o r o s p h e r e s y o u u s e to p e r fo r m a lig nm e nt v e r ific a tio n de p e nd u p o n w h ic h a p p lic a tio n y o u a r e u s ing , a s s h o w n in th e c h a r t b e lo w . F o llo w th e p a c k a g e ins e r t ins tr u c tio ns fo r m ix ing a nd h a ndling flu o r o s p h e r e s .

Ap p lica tion Flow -Ch eck Flow -Ch eck 770 Flow -Ch eck 675

FS -S S -FIT C-PE -E CD -PC5 c

FS -S S -FIT C-PE -E CD -PC5-PC7 c c

FS -S S -FIT C-PE -E CD -APC c c

FS -S S -FIT C-PE -E CD -APC-PC7 c c c

PN 624923B 3-3

QUALITY CONTROLD A IL Y Q C 3

2 P r e p a r e th e a p p r o p r ia te F lo w -S e t flu o r o s p h e r e s fo r th e a p p lic a tio ns y o u ne e d to r u n. T h e

F lo w - S e t flu o r o s p h e r e s tu b e is u s e d to s e t th e de te c to r g a ins a nd v o lta g e s to th e r e q u ir e d le v e l. T h e flu o r o s p h e r e s y o u u s e to p e r fo r m s ta nda rdiz a tio n de p e nd u p o n w h ic h a p p lic a tio n y o u a r e u s ing , a s s h o w n in th e c h a r t b e lo w . F o llo w th e p a c k a g e ins e r t ins tr u c tio ns fo r m ix ing a nd h a ndling th e s e flu o r o s p h e r e s .

3 P r e p a r e A u to S e tu p c o m p e ns a tio n tu b e s

fo r e a c h flu o r o c h r o m e in th e a p p lic a tio n y o u ne e d to r u n.

S ta in w ith a p p r o p r ia te s ing le c o lo r r e a g e nts , s u c h a s CD 45, in e a c h flu o r o c h r o m e .

4 P r e p a r e a n A u to S e tu p v e r ific a tio n tu b e .

a . U s e th e a p p r o p r ia te a m o u nt o f Im m u no -T r o l Ce lls o r I m m u no -T r o l Lo w Ce lls a c c o rding to th e ins tr u c tio ns o n th e p a c k a g e ins e r t o r r e c o ns titu te th e Cy to -tr o l Co ntr o l Ce lls a c c o rding to th e ins tr u c tio ns o n th e p a c k a g e ins e r t.

b . S ta in th e c e lls w ith th e m o no c lo na l a ntib o die s y o u u s e fo r th e p r o to c o l o r p a ne l.

5 R u n th e A u to S e tu p S c h e du le r.

Ap p lica tion Flow -Set Flow -Set 770 Flow -Set 675

FS -S S -FIT C-PE -E CD -PC5 c

FS -S S -FIT C-PE -E CD -PC5-PC7 c c

FS -S S -FIT C-PE -E CD -APC c c

FS -S S -FIT C-PE -E CD -APC-PC7 c c c

PN 624923B3-4

QUALITY CONTROLA U TOSETU P SCH ED U L ER

3.4 AUTOSETUP SCHEDULERU s e th e A u to S e tu p S c h e du le r to s e le c t th e a p p lic a tio ns th a t y o u ne e d to r u n o n th e ins tr u m e nt in a g iv e n da y o r " s h ift" . A u to S e tu p S c h e du le r g r o u p s th e s e le c te d a p p lic a tio ns a nd p r o v ide s th e c a r o u s e l lo a d r e p o r t to fa c ilita te s e tting u p a nd lo a ding s a m p le s fo r da ily Q C.

1 Tools tt AutoSetup Sch ed uler.

2 th e a p p lic a tio ns y o u w a nt to

s c h e du le fo r a g iv e n da y o r s h ift.

If y o u w a nt to s c h e du le a n a p p lic a tio n th a t do e s no t a p p e a r in th e lis t, u s e th e A p p lic a tio n D e finitio n W iz a rd to de fine th e a p p lic a tio n.

N ote: U s ing th e de s c r ip to r Q C in fr o nt o f a ny p r o to c o l na m e a p p e nds th e r e s u lts o f m u ltip le F lo w -Ch e c k r u ns to a s ing le s p r e a ds h e e t. S e e W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns to a u to m a tic a lly e x p o r t da ta to MS E x c e l fo r Q u a lity Co ntr o l m o nito r ing o r b a s ic s p r e a ds h e e t da ta e ntr y .

3 E nte r th e Ca r o u s e l nu m b e r a nd .

PN 624923B 3-5

QUALITY CONTROLA U TOSETU P II W IZ A R D 3

3.5 AUTOSETUP II WIZ ARDA u to S e tu p I I s im p lifie s a nd a u to m a te s a p p lic a tio n s e tu p a nd Q C m o nito r ing fo r m u ltip le a p p lic a tio ns s im u lta ne o u s ly . Y o u c a n p e r fo r m a Q C s e tu p fo r a ll o f y o u r m u ltic o lo r a p p lic a tio ns th a t y o u r u n du r ing a s ing le da y o r s h ift a t th e s a m e tim e fo r c o m m o n ins tr u m e nt o p tic a l c o nfig u r a tio ns u s ing th e A p p lic a tio n D e finitio n W iz a rd a nd th e A u to S e tu p S c h e du le r.

Before Running th e AutoSetup II Wiz a rd

r Ch e c k th a t th e D A ILY S T A R T U P p r o c e du r e w a s p e r fo r m e d.

r A llo w a b o u t 40 m inu te s to w a r m u p th e s y s te m .

r Ch e c k th e Cy to m e te r s ta tu s m e s s a g e . S ta r t th is p r o c e du r e w h e n th e Cy to m e te r s ta tu s m e s s a g e A waiting S ample a p p e a r s . R e c o rd a ny e r r o r m e s s a g e s in y o u r ins tr u m e nt’s lo g b o o k .

r S e t u p W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns to p r int r e s u lts .

r S e t u p W o r k s p a c e P r e fe r e nc e s - LMD F ile N a m e to s p e c ify lis tm o de file na m e s .

4 to p r int th e Ca r o u s e l

Lo a d R e p o r t.

N ote: I f a b s o lu te c o u nts a r e r e q u ir e d,

a nd u s e th e A c q u is itio n Ma na g e r to e nte r th e CA L F a c to r.

5 P la c e tu b e s in th e c a r o u s e l in th e o rde r

s h o w n o n th e Ca r o u s e l Lo a d R e p o r t.

o n th e Ca r o u s e l Lo a d R e p o r t s c r e e n to r u n A u to S e tu p II .


r D aily S tartup .



PN 624923B3-6

QUALITY CONTROLA U TOSETU P II W IZ A R D

r F o llo w th e dir e c tio ns in th e F lo w -Ch e c k flu o r o s p h e r e s p a c k a g e ins e r t to e s ta b lis h e x p e c te d r a ng e v a lu e s fo r y o u r la b o r a to r y .

r E s ta b lis h F lo w - S e t flu o r o s p h e r e s ta r g e t c h a nne ls fo r y o u r la b o r a to r y a nd m o dify th e de fa u lt a p p lic a tio n de finitio ns if ne c e s s a r y . F o llo w th e dir e c tio ns in th e F lo w - S e t flu o r o s p h e r e s p a c k a g e ins e r t.

r S e t u p th e Q C P r o du c ts th a t y o u ne e d to u s e w ith A u to S e tu p a p p lic a tio ns .

r S e t u p r e g io n s ta tis tic s e x p o r ting a nd a s s ig n Q C p r o du c ts to r e g io ns .

r If a c q u is itio n e nds a s a r e s u lt o f du r a tio n (no t s to p c o u nt) w h ile r u nning A u to S e tu p II ,

s e le c t o n th e Cy to m e te r Co ntr o l to o lb a r to c o ntinu e .

r B e fo r e u s ing a ne g a tiv e c o ntr o l s a m p le w ith A u to S e tu p , y o u m u s t Mo dify A S ta nda rdiz a tio n P r o to c o l fo r u s e in th e A u to S e tu p p a ne l. S e e U s ing N e g a tiv e Co ntr o ls w ith A u to S e tu p in th e R e fe r e nc e Ma nu a l.

1 o n th e Ca r o u s e l Lo a d

R e p o r t s c r e e n in th e A u to S e tu p S c h e du le r.

2 A fte r da ta a c q u is itio n s ta r ts v ie w a ll

h is to g r a m s to e ns u r e p e a k s fa ll w ith in r e g io ns .

a . S e tu p Mo de o n th e Cy to m e te r Co ntr o l A c q u is itio n S e tu p T a b .

b . A dju s t v o lta g e s to p la c e p e a k s w ith in r e g io ns

c . U nc h e c k S e tu p Mo de .

PN 624923B 3-7


N ote: W h e n r u nning a ne g a tiv e c o ntr o l v e r ify th a t th e A u to G a te is a r o u nd th e p o p u la tio n o f inte r e s t in th e F S v s S S do tp lo t. If y o u ne e d to , a dju s t p o p u la tio ns .

a . Auto Ad just Disa b led a s s o o n a s th e A u to S e tu p w iz a rd dis p la y s .

b . a dju s t v o lta g e s to p la c e th e ly m p h o c y te s in a s u ita b le p o s itio n.

c . a dju s t v o lta g e s to p la c e th e flu o r e s c e nc e p o p u la tio ns w ith in th e fir s t lo g de c a de .

d. .

T h e W iz a rd a dju s ts V o lts a nd G a ins to p la c e th e F lo w -S e t flu o r o s p h e r e s w ith in th e r e g io ns . T h e W iz a rd dia lo g b o x dis p la y s a lis t o f th e p a r a m e te r s r e m a ining to b e a dju s te d. If th e W iz a rd c a nno t a dju s t p e a k s to th e r e g io ns w ith in 60 s e c o nds , A u to S e tu p a b o r ts a nd th e p a r a m e te r s th a t fa ile d a r e lis te d.

T h e c o m p e ns a tio n tu b e s a r e u s e d to c a lc u la te th e c o m p e ns a tio n c o e ffic ie nts r e q u ir e d, a ll th e s e tu b e s a r e r u n in th e s a m e w a y .

PN 624923B3-8

QUALITY CONTROLA U TOSETU P II W IZ A R D

T h is p r o c e s s is r e p e a te d fo r e a c h o f th e c o m p e ns a tio n tu b e s ( fo r e x a m p le : F L2, F L3, F L4, a nd F L5) .

V e r ific a tio n s a m p le s a r e r u n to e ns u r e th e c a lc u la te d c o e ffic ie nts a r e s a tis fa c to r y fo r th e s a m p le s a nd a ntib o die s y o u a r e u s ing in y o u r te s ts .

N ote: I f m u ltip le a p p lic a tio ns a r e s c h e du le d to g e th e r, s a m p le s c o m m o n to th e a p p lic a tio ns a r e p r o c e s s e d in ta nde m w ith o u t dr o p p ing th e tu b e . T h e p r o to c o l is dis p la y e d du r ing p r o c e s s ing .

3 to p r int v e r ific a tio n

tu b e r e s u lts a nd a dv a nc e to th e a p p r o v e s c r e e n.

PN 624923B 3-9


4 Ch e c k th e p r inte d v e r ific a tio n tu b e

r e s u lts to e ns u r e ins tr u m e nt s e tting s

a r e c o r r e c t a nd .

N ote: D u e to diffe r e nc e s b e tw e e n indiv idu a l ins tr u m e nts , s e tting s file s s h o u ld no t b e tr a ns fe r r e d fr o m o ne ins tr u m e nt to a no th e r. I f y o u a tte m p t to u s e s e tting s file s fr o m a no th e r ins tr u m e nt, th e s o ftw a r e dis p la y s Incorrect C y tometer S erial N umber.

5 R e v ie w Q C R e s u lts in th e Q C D a ta b a s e .

PN 624923B3-1 0

QUALITY CONTROLA P P L ICA TION D EF INITION W IZ A R D

3.6 APPLICATION DEFINITION WIZ ARDU s e th e A p p lic a tio n D e finitio n W iz a rd to de fine y o u r a p p lic a tio ns a nd s a v e th e de finitio ns fo r u s e b y th e A u to S e tu p S c h e du le r. T h e a p p lic a tio n de finitio n c a p tu r e s th e ins tr u m e nt s e tu p , la s e r s , p a r a m e te r s , flu o r o c h r o m e s , ta r g e t c h a nne ls , v e r ific a tio n a nd a lig nm e nt r e q u ir e m e nts o f a p a r tic u la r a p p lic a tio n.

Before Running th e Ap p lica tion Definition Wiz a rd

r E s ta b lis h F lo w - S e t flu o r o s p h e r e s ta r g e t c h a nne ls fo r y o u r la b o r a to r y . F o llo w th e dir e c tio ns in th e F lo w -S e t flu o r o s p h e r e s p a c k a g e ins e r t.

r E ns u r e th a t th e b a s e p r o to c o l y o u u s e to de fine a n a p p lic a tio n c o nta ins o nly th e p a r a m e te r s th a t y o u u s e in th e a p p lic a tio n. If it c o nta ins e x tr a u nu s e d p a r a m e te r s , th e c o m p e ns a tio n m a tr ix w ill b e inc o r r e c t.

1 Tools tt AutoSetup Ap p lica tion

Definition.

2 o ne o f th e fo llo w ing o p tio ns :

Ap p lica tion a v a ila b le only to current userAp p lica tion a v a ila b le to a ll users (com m on).

3 o ne o f th e fo llo w ing o p tio ns :

Crea te a new a p p lica tion d efinitionMod ify a n a p p lica tion d efinition

a nd th e n .

PN 624923B 3-1 1

QUALITY CONTROLA P P L ICA TION D EF INITION W IZ A R D 3

4 a nd s e le c t th e p r o to c o l

to u s e a s th e b a s e p r o to c o l fo r th e a p p lic a tio n.

T h e P r o to c o l S u m m a r y dis p la y s a de s c r ip tio n o f th e p a r a m e te r s a nd la s e r s e tu p fo r th e s e le c te d p r o to c o l.

.

5 th e filte r b lo c k to u s e fo r th e

a p p lic a tio n a nd .

N ote: A n a lig nm e nt a p p lic a tio n m a y no t h a v e a U s e r de fine d F ilte r B lo c k s e le c te d. A lig nm e nt a p p lic a tio ns m u s t h a v e e ith e r a 1-La s e r F ilte r B lo c k o r 2-La s e r F ilte r B lo c k s e le c te d.

6 th e fo r w a rd s c a tte r c o lle c tio n

a ng le to u s e fo r th e a p p lic a tio n a nd

.

PN 624923B3-1 2


7 th e dy e s y o u u s e fo r e a c h

flu o r e s c e nc e de te c to r a nd

.

8 S e le c t a Q C p r o du c t a nd e nte r a ta r g e t

c h a nne l fo r F o r w a rd S c a tte r, S ide S c a tte r a nd e a c h flu o r e s c e nc e de te c to r.

N ote: I f y o u a r e r u nning a 5 c o lo r 2 la s e r a p p lic a tio n, e nte r F lu o r e s c e nc e a nd F o r w a rd S c a tte r ta r g e t c h a nne ls fo r th e F lo w -S e t 67 5 a nd F lo w - S e t 7 7 0 F lu o r o s p h e r e s .

PN 624923B 3-1 3


9 E nte r na m e s fo r th e a p p lic a tio n a nd th e

ins tr u m e nt s e tting s file a nd

.

10 Ch o o s e to h a v e th e w iz a rd c r e a te a

v e r ific a tio n p r o to c o l o r a dd a s a v e d v e r ific a tio n p r o to c o l o r p a ne l. U s e th e b u tto ns to a dd, r e m o v e a nd r e o rde r m u ltip le v e r ific a tio n p r o to c o ls .

.

N ote: A ny p a ne ls u s e d fo r Q u a lity

Co ntr o l m u s t b e c o ns tr u c te d w ith u niq u e indiv idu a l p r o to c o ls .

11 A s u m m a r y r e p o r t is dis p la y e d fo r th e

a p p lic a tio n y o u ju s t de fine d. T h e a p p lic a tio n de finitio n is a v a ila b le fo r u s e w ith th e A u to S e tu p S c h e du le r.

to p r int th e s u m m a r y

r e p o r t a nd to e x it th e w iz a rd.

PN 624923B3-1 4


Setting Up QC Prod ucts

Ex p orting Reg ion Results To QC Da ta b a se a nd Assig ning QC Prod ucts

1 to dis p la y

th e R e p o r t G e ne r a to r to o lb a r.

2 o n th e R e p o r t G e ne r a to r

to o lb a r to dis p la y th e E dit P r o du c ts s c r e e n.

3 R e fe r to th e p a c k a g e ins e r t a nd e nte r

info r m a tio n a b o u t th e Q C P r o du c t in th e a p p r o p r ia te fie lds .

4 .

1 o n a r e g io n to m a k e it a c tiv e .

2 R ig h t m o u s e c lic k o n th e r e g io n a nd

Reg ion Prop erties.

PN 624923B 3-1 5


Rev iew QC Results

3 E ns u r e th a t Reg ion Sta tistics ex p orted

for Qua lity Control.

4 ne x t to QC Prod uct a nd s e le c t

th e Q C P r o du c t to a s s ig n to th e r e g io n.

5 a nd s a v e th e p r o to c o l.

1 to dis p la y

th e R e p o r t G e ne r a to r to o lb a r.

2 o n th e R e p o r t G e ne r a to r

to o lb a r to dis p la y th e Q C Le v e y Je nning s s c r e e n.

PN 624923B3-1 6


3 ne x t to Ap p lica tion

a nd to s e le c t a n a p p lic a tio n.

4 Tem p la te tt Op en Tem p la te.

5 th e de s ir e d te m p la te a nd

.

6 R e p e a t s te p s 3 - 5 to v ie w o th e r Q C da ta p lo ts .

S e e Q C Le v e y Je nning s S c r e e n in th e R e fe r e nc e m a nu a l fo r info r m a tio n a b o u t c r e a ting a nd e diting Q C p lo ts .

PN 624923B 4-1

44RUNNING SAMPLES

4.1 SAMPLE REQUIREMENTSA t le a s t 0 .5 m L o f p r e p a r e d s a m p le is ne e de d. I t m u s t b e in a 12- x 7 5-m m te s t tu b e . S a m p le s a na ly z e d o n th e ins tr u m e nt m u s t b e in a s ing le -c e ll s u s p e ns io n. T y p ic a lly , c e lls a r e p r e p a r e d b e fo r e th e y a r e a na ly z e d. T h e m e th o d u s e d to p r e p a r e a s p e c im e n de p e nds o n th e s a m p le ty p e a nd th e a s s a y de s ir e d. F o r e x a m p le , a T Q -P r e p w o r k s ta tio n c o m b ine d w ith a P r e p P lu s o r P r e p P lu s 2 le ts y o u p r e p a r e a ntib o dy -la b e le d c e lls fr o m a n a ntic o a g u la te d w h o le - b lo o d s p e c im e n fo r s u r fa c e m a r k e r a na ly s is .

In g e ne r a l, th e o p tim u m c o nc e ntr a tio n fo r a na ly s is is 5 x 10 6 c e lls /m L. W h e n th is c o nc e ntr a tio n is no t p o s s ib le , r e fe r to th e p a c k a g e ins e r t fo r th e p r e p a r a tio n m e th o d y o u a r e u s ing .

T h e ins tr u m e nt c a n m e a s u r e c e lls th a t a r e b e tw e e n 0 .5 µ m a nd 40 µ m in dia m e te r.

4.2 BEFORE RUNNING SAMPLES

1 Ch e c k th a t th e D A ILY S T A R T U P p r o c e du r e s w e r e do ne .

2 Ch e c k th a t th e D A ILY Q C p r o c e du r e s w e r e do ne .

3 E ns u r e A c q u is itio n O p tio ns a nd LMD file na m e a r e s e t u p in W o r k s p a c e P r e fe r e nc e s .

4 E ns u r e th e r e is s u ffic ie nt s p a c e o n y o u r h a rd dr iv e fo r s a m p le p r o c e s s ing a nd da ta

a c q u is itio n.

5 R e a d th e I m p o r ta nt Info r m a tio n o n th e S u p p o r t ta b in th e R e s o u rc e E x p lo r e r .


r D aily S tartup .



PN 624923B4-2

RUNNING SAMPLESR U NNING SA M P L ES - M CL A U TOM A TIC M OD E

4.3 RUNNING SAMPLES - MCL AUTOMATIC MODESHOW ME a n e x a m p le o f MCL A u to m a tic Mo de s a m p le p r o c e s s ing .

IMPORTANT R is k of s amp le mis id entification if a p ow er failure occurs d uring s amp le p roces s ing . In th e event of a p ow er failure, d is card any in-p roces s s amp les .

1 P r e p a r e s a m p le s a c c o rding to th e

r e a g e nt p a c k a g e ins e r t.

2 P la c e th e s a m p le tu b e s in a c a r o u s e l.

CAUTION Pos s ib le flow cell d amag e. T o avoid clog g ing th e s amp le p rob e, s amp le tub ing or flow cell, ens ure th at 1 2 mm x 7 5 mm tes t tub es are free of d eb ris b efore y ou us e th em.

PN 624923B 4-3

RUNNING SAMPLESR U NNING SA M P L ES - M CL A U TOM A TIC M OD E 4

3 O p e n th e MCL c o v e r a nd p la c e th e

c a r o u s e l o n th e MCL.


WARNING R is k of injury . D o not op en th e M CL cover w h ile th e M CL is moving . T o avoid injury , w ait until th e M CL s top s moving b efore op ening th e M CL cover.

PN 624923B4-4

RUNNING SAMPLESR U NNING SA M P L ES - M CL A U TOM A TIC M OD E

5 U s e th e A c q u is itio n Ma na g e r T o o lb a r

o r D r a g A nd D r o p to s e le c t th e W o r k lis t, P a ne l o r P r o to c o l y o u w a nt to u s e .

N ote: I f y o u lo a d a P a ne l th a t c o nta ins a de le te d P r o to c o l, th e s o ftw a r e p r o m p ts y o u to u s e th e N e w P a ne l W iz a rd to c o r r e c t th e p r o b le m .

6 E nte r th e c a r o u s e l ID nu m b e r in th e

A c q u is itio n Ma na g e r.

7 to b e g in p r o c e s s ing s a m p le s .

D u r ing th e s a m p le c y c le th e fo llo w ing s e r ie s o f Cy to m e te r s ta tu s m e s s a g e s a p p e a r s :

A waiting S ample

P reparing S ample

A cquiring

S topping.

8 O b s e r v e th e E v e nts /S e c c o u nte r o n th e S ta tu s B a r to m o nito r da ta a c q u is itio n.

N ote: W h ile r u nning a W o r k lis t a nd u s ing b a r c o de s o n y o u r s a m p le tu b e s , if th e T u b e ID in th e W o r k lis t do e s no t m a tc h th e b a r c o de o n th e tu b e , a s y s te m m e s s a g e a p p e a r s . A fte r y o u a c k no w le dg e th e m e s s a g e , th e m is m a tc h e d tu b e is a b o r te d a nd th e p r o c e s s ing s to p s fo r th e r e s t o f th e c a r o u s e l.

IMPORTANT R is k of erroneous res ults if th e s amp le b ecomes d iluted . W h en op erating th e Cy tometer, if th e M CL cover is op ened , an aud ib le alarm s ound s and a w arning mes s ag e is d is p lay ed . If acq uis ition comp letes w h ile th e M CL cover is op ened , d o not us e th e s amp le for s ub s eq uent p roces s ing . U nd er th is cond ition, th e current s amp le may b e d iluted at th e end of acq uis ition.

PN 624923B 4-5

RUNNING SAMPLESR U NNING SA M P L ES - M CL M A NU A L M OD E 4

4.4 RUNNING SAMPLES - MCL MANUAL MODEMCL Ma nu a l Mo de o p e r a te s th e s a m e a s MCL A u to m a tic Mo de e x c e p t th e MCL s to p s a fte r p r o c e s s ing e a c h tu b e to a llo w y o u to e dit th e S a m p le ID .

.

9 V e r ify a ll s a m p le ID s b e fo r e r e p o r ting r e s u lts .

10 P r int P a tie nt P a ne l R e p o r t, if ne e de d. S e e P a ne l R e p o r t in th e R e fe r e nc e m a nu a l.

IMPORTANT R is k of rep orting incorrect res ults . D ata d is p lay s for lig h t s catter p atterns , antib od y s taining p rofiles , and all g ates and b ound aries us ed to arrive at th e tes t res ult s h ould b e review ed b y a lab oratory p rofes s ional w h en interp reting th e d ata.



2 P la c e th e s a m p le tu b e s in a c a r o u s e l.


PN 624923B4-6

RUNNING SAMPLESR U NNING SA M P L ES - M CL M A NU A L M OD E

3 O p e n th e MCL c o v e r a nd p la c e th e

c a r o u s e l o n th e MCL.


5 T o s to p a fte r p r o c e s s ing e a c h tu b e a nd

e dit th e S a m p le ID ,

Ed it Sa m p le IDs o n W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns .


PN 624923B 4-7

RUNNING SAMPLESR U NNING SA M P L ES - M CL M A NU A L M OD E 4




7 E nte r th e c a r o u s e l ID nu m b e r in th e

A c q u is itio n Ma na g e r.

8 to p la c e th e Cy to m e te r in MCL Ma nu a l Mo de .

9 to b e g in p r o c e s s ing s a m p le s .


PN 624923B4-8

RUNNING SAMPLESR U NNING SA M P L ES - M CL M A NU A L M OD E

10 E nte r th e tu b e lo c a tio n to p r o c e s s a nd

.


A waiting S ample

P reparing S ample

A cquiring

S topping.


N ote: W h ile r u nning a W o r k lis t, if th e T u b e I D in th e W o r k lis t do e s no t m a tc h th e b a r c o de o n th e tu b e , a s y s te m m e s s a g e a p p e a r s . A fte r y o u a c k no w le dg e th e m e s s a g e , th e m is m a tc h e d tu b e is a b o r te d a nd th e p r o c e s s ing s to p s fo r th e r e s t o f th e c a r o u s e l.

12 If y o u e na b le d Ed it Sa m p le IDs o n

W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns , e dit th e s a m p le I D a fte r th e tu b e finis h e s p r o c e s s ing a nd

.

N ote: A fte r th e fir s t tu b e in a p a ne l, S a m p le ID 1 c a nno t b e c h a ng e d.

13 E nte r th e lo c a tio n o f th e ne x t tu b e to

p r o c e s s a nd .

14 R e p e a t s te p s 12 a nd 13 to p r o c e s s a dditio na l tu b e s .

PN 624923B 4-9

RUNNING SAMPLESR U NNING SA M P L ES - SING L E TU B E M OD E 4

4.5 RUNNING SAMPLES - SINGLE TUBE MODET h e S ing le T u b e Mo de o p e r a te s b y intr o du c ing tu b e s indiv idu a lly into th e MCL th r o u g h th e MCL T u b e A c c e s s do o r. SHOW ME a n e x a m p le o f S ing le T u b e Mo de s a m p le p r o c e s s ing .

N ote: W h e n o p e r a ting in th e S ing le T u b e Mo de , th e P r im e fu nc tio n is no t a v a ila b le w h e n th e s y s te m is b e tw e e n s a m p le s , e v e n th o u g h th e to o lb a r is a c tiv e .

.






2 If y o u w a nt th e MCL to s to p a fte r

p r o c e s s ing e a c h tu b e to a llo w y o u to

e dit th e S a m p le ID , Ed it

Sa m p le IDs o n W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns .


PN 624923B4-1 0

RUNNING SAMPLESR U NNING SA M P L ES - SING L E TU B E M OD E




4 to p la c e th e Cy to m e te r in S ing le T u b e Mo de . T h e MCL m o v e s to p u t c a r o u s e l

p o s itio n 10 u nde r th e T u b e A c c e s s do o r.

5 O p e n th e MCL T u b e A c c e s s do o r a nd

p la c e th e s a m p le tu b e in c a r o u s e l p o s itio n 10 .



PN 624923B 4-1 1

RUNNING SAMPLESR U NNING SA M P L ES - SING L E TU B E M OD E 4

6 Clo s e th e MCL T u b e A c c e s s do o r a nd

to b e g in p r o c e s s ing s a m p le s .


A waiting S ample

P reparing S ample

A cquiring

S topping.


N ote: W h ile r u nning a W o r k lis t, if th e T u b e I D in th e W o r k lis t do e s no t m a tc h th e b a r c o de o n th e tu b e , a s y s te m m e s s a g e a p p e a r s . A fte r y o u a c k no w le dg e th e m e s s a g e , th e m is m a tc h e d tu b e is a b o r te d a nd th e p r o c e s s ing s to p s fo r th e r e s t o f th e c a r o u s e l.


9 If y o u e na b le d Ed it Sa m p le IDs o n

W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns , e dit th e s a m p le ID a nd

.

N ote: A fte r th e fir s t tu b e in a p a ne l, S a m p le ID 1 c a nno t b e c h a ng e d.


PN 624923B4-1 2

RUNNING SAMPLESR U NNING SA M P L ES - SING L E TU B E M OD E

10 R e p e a t s te p s 5 th r o u g h 9 to p r o c e s s a dditio na l tu b e s .

to e x it S ing le T u b e Mo de a nd r e tu rn to A u to m a tic Mo de .


PN 624923B 5-1

55OPERATION PRINCIPLES

5.1 WHAT THIS CHAPTER EX PLAINST h is c h a p te r e x p la ins h o w th e Cy to m e te r m e a s u r e s s c a tte r e d lig h t a nd flu o r e s c e nc e a s c e lls p a s s th r o u g h th e la s e r b e a m .

T h e illu s tr a tio ns in th is c h a p te r a r e no t e x a c t r e p r e s e nta tio ns o f th e ins ide o f th e Cy to m e te r. T h e y a r e fo r e x p la na to r y p u r p o s e s o nly .

5.2 SAMPLE FLOW

Sa m p le Loa d ing

T h e s a m p le c a r o u s e l h a s b a r- c o de la b e ls th a t ide ntify th e c a r o u s e l a nd th e tu b e p o s itio n nu m b e r. A ls o , y o u c a n p u t b a r-c o de la b e ls o n th e s a m p le tu b e s . S e e A p p e ndix B , B A R -CO D E S P E CIF ICA T IO N S .

T h e MCL h a s a b a r- c o de r e a de r th a t r e a ds th e c a r o u s e l nu m b e r, th e s a m p le tu b e p o s itio n, a nd th e s a m p le tu b e b a r-c o de la b e ls a s th e c a r o u s e l r o ta te s . T h e MCL h a ndle s a s a m p le tu b e a s fo llo w s :

r It lifts th e tu b e o u t o f th e c a r o u s e l into a c e nte r ing c u p .

r It m o v e s th e b o tto m o f th e tu b e in a c irc u la r o r b it fo r 3 s e c o nds to m ix th e s a m p le .

r It lo w e r s its s a m p le p r o b e into th e tu b e a nd th e tu b e is p r e s s u r iz e d. S a m p le flo w b e g ins .

T h e s a m p le p r o b e is c le a ne d a u to m a tic a lly w h e n s a m p le flo w e nds .

Hy d rod y na m ic Focusing

If th e c e lls w e r e to m o v e th r o u g h th e la s e r b e a m in diffe r e nt w a y s du r ing s a m p le flo w , s a m p le a na ly s is c o u ld b e dis to r te d. T h e ins tr u m e nt u s e s a p r o c e s s c a lle d h y dr o dy na m ic fo c u s ing to e ns u r e th a t th e c e lls m o v e th r o u g h th e la s e r b e a m o ne a t a tim e , a lo ng th e s a m e p a th . Hy dr o dy na m ic fo c u s ing o c c u r s in th e flo w c e ll.

T h e flo w c e ll ( F ig u r e 5.1) c o nta ins a r e c ta ng u la r c h a nne l. A p r e s s u r iz e d s tr e a m o f s h e a th flu id e nte r s th e c h a nne l a t th e lo w e r e nd a nd flo w s u p w a rd. T h e s e ns ing a r e a o f th e flo w c e ll is a t th e c e nte r o f th e c h a nne l.

W h ile th e s h e a th s tr e a m is flo w ing th r o u g h th e c h a nne l, a s tr e a m o f s a m p le is inje c te d into th e m iddle o f th e s h e a th s tr e a m . A s s h o w n in F ig u r e 5.1, th e s h e a th s tr e a m s u r r o u nds , b u t do e s no t m ix w ith , th e s a m p le s tr e a m . T h e p r e s s u r e o f th e s h e a th s tr e a m fo c u s e s th e s a m p le s tr e a m s o th a t th e c e lls flo w th r o u g h th e la s e r b e a m s ing le file .


PN 624923B5-2

OPERATION PRINCIPLESSA M P L E F L OW

Fig ure 5.1 Flow Cell (Hy d rod y na m ic Focusing )

b T o w a s te g T o w a s te

c S e ns ing a r e a h a s r e c ta ng u la r c h a nne l h S a m p le s tr e a m e nte r s h e r e

d S a m p le s tr e a m i S h e a th s tr e a m e nte r s h e r e

e S h e a th s tr e a m j D ir e c tio n o f la s e r b e a m

f Ce lls

b

c

de

f

gi

h

j

PN 624923B 5-3

OPERATION PRINCIPLESL A SER B EA M SH A P ING 5

5.3 LASER BEAM SHAPINGB e fo r e th e la s e r b e a m r e a c h e s th e s a m p le s tr e a m , c r o s s - c y lindr ic a l le ns e s fo c u s th e b e a m ( s e e F ig u r e 5.2) . F o c u s ing k e e p s th e b e a m p e r p e ndic u la r to th e s a m p le s tr e a m flo w w h ile m a k ing th e b e a m s m a ll e no u g h to illu m ina te o nly o ne c e ll a t a tim e .

T h e fir s t le ns c o ntr o ls th e w idth o f th e b e a m ; th e s e c o nd, th e h e ig h t. T h e r e s u lting e llip tic a l b e a m is fo c u s e d o n th e s e ns ing a r e a o f th e flo w c e ll.

Fig ure 5.2 La ser Bea m Sh a p ing

b Cro s s s e c tio n b e fo r e le ns e s g V e r tic a l b e a m s h a p ing le ns

c Cro s s s e c tio n b e fo r e v e r tic a l le ns h La s e r b e a m dir e c tio n

d Cro s s s e c tio n b e fo r e h o r iz o nta l le ns i Ho r iz o nta l b e a m s h a p ing le ns

e T o w a s te j La s e r b e a m

f F lo w c e ll

g

i

j

b

c

d

e

f

h

PN 624923B5-4

OPERATION PRINCIPLESCEL L IL L U M INA TION

5.4 CELL ILLUMINATIONA s c e lls in th e s a m p le s tr e a m g o th r o u g h th e s e ns ing a r e a o f th e flo w c e ll, th e e llip tic a l la s e r b e a m illu m ina te s th e m . T h e c e lls s c a tte r th e la s e r lig h t a nd e m it flu o r e s c e nt lig h t fr o m flu o r e s c e nt dy e s a tta c h e d to th e m .

Forw a rd Sca tter

T h e a m o u nt o f la s e r lig h t s c a tte r e d a t na r r o w a ng le s to th e a x is o f th e la s e r b e a m is c a lle d fo r w a rd s c a tte r ( F S ) . T h e a m o u nt o f F S is p r o p o r tio na l to th e s iz e o f th e c e ll th a t s c a tte r e d th e la s e r lig h t.

Sid e Sca tter a nd Fluorescent Lig h t

T h e a m o u nt o f la s e r lig h t s c a tte r e d a t a b o u t a 90 ° a ng le to th e a x is o f th e la s e r b e a m is c a lle d s ide s c a tte r ( S S ) . T h e a m o u nt o f S S is p r o p o r tio na l to th e g r a nu la r ity o f th e c e ll th a t s c a tte r e d th e la s e r lig h t. F o r e x a m p le , S S is u s e d to diffe r e ntia te b e tw e e n ly m p h o c y te s , m o no c y te s , a nd g r a nu lo c y te s .

In a dditio n to th e S S , th e c e lls e m it flu o r e s c e nt lig h t ( F L) a t a ll a ng le s to th e a x is o f th e la s e r b e a m . T h e a m o u nt o f F L e na b le s th e ins tr u m e nt to m e a s u r e c h a r a c te r is tic s o f th e c e lls e m itting th a t lig h t, de p e nding o n th e r e a g e nts u s e d. F o r e x a m p le , F L is u s e d to ide ntify m o le c u le s , s u c h a s c e ll s u r fa c e a ntig e ns .

5.5 LIGHT COLLECTION, SEPARATION AND MEASUREMENT

Forw a rd Sca tter Collection

T h e F S s e ns o r ( s e e F ig u r e 5.3 a nd F ig u r e 5.4) c o lle c ts th e fo r w a rd s c a tte r — th e la s e r lig h t th a t is s c a tte r e d a t na r r o w a ng le s to th e a x is o f th e la s e r b e a m . W h e n lig h t r e a c h e s th e F S s e ns o r, th e s e ns o r g e ne r a te s v o lta g e p u ls e s ig na ls . T h e s e s ig na ls a r e p r o p o r tio na l to th e a m o u nt o f lig h t th e s e ns o r r e c e iv e s . A s e x p la ine d in He a ding 5.6, S IG N A L P R O CE S S IN G , th e s ig na ls a r e p r o c e s s e d to m e a s u r e th e c h a r a c te r is tic s o f th e c e lls th a t s c a tte r e d th e lig h t.

T h e F S s e ns o r c o nta ins a F ie ld S to p th a t a llo w s y o u to c o lle c t diffe r e nt a ng le s o f F o r w a rd S c a tte r. T h e s ta nda rd ( fa c to r y s e t) p o s itio n c o lle c ts F S a ng le s o f 1 to 19° . T h e s e c o nd p o s itio n a llo w s y o u to c o lle c t a lo w e r a ng le o f F S , 1 to 8 ° . S e e P O S I T IO N T HE F I E LD S T O P .

PN 624923B 5-5

OPERATION PRINCIPLESL IG H T COL L ECTION, SEP A R A TION A ND M EA SU R EM ENT 5

Fig ure 5.3 Op tica l Sy stem w ith Op tiona l Red Solid -sta te La ser

Fig ure 5.4 Op tica l Sy stem w ith Op tiona l HeNe La ser

b Arg on (488) las er f FL 4 j Field s top

c S S s ens or g FL 5 1) FS s ens or

d S olid -s tate (635) las er (op tional) h FL 2 1! Flow cell

e FL 3 i FL 1

b Arg on (488) las er f FL 4 j Field s top

c S S s ens or g FL 5 1) FS s ens or

d H eNe (633) las er (op tional) h FL 2 1! Flow cell

e FL 3 i FL 1

b c d

e

f

g

hij

1)1!

b c

d

e

f

g

hij

1)1!

PN 624923B5-6

OPERATION PRINCIPLESL IG H T COL L ECTION, SEP A R A TION A ND M EA SU R EM ENT

Sid e Sca tter a nd Fluorescent Lig h t Collection

In o rde r fo r th e s e ns o r s to m e a s u r e S S a nd F L, th e lig h t m u s t b e c o lle c te d a nd th e S S a nd flu o r e s c e nt lig h t m u s t b e s e p a r a te d.

T h e p ic k u p le ns /s p a tia l filte r a s s e m b ly c o lle c ts S S a nd F L fr o m o nly th e s e ns ing a r e a o f th e flo w c e ll, a nd c o llim a te s it. T h is lig h t th e n g o e s to w a rd th e S S s e ns o r.

Sid e Sca tter

T h e w a v e le ng th o f S S is 48 8 nm . It is m u c h m o r e inte ns e th a n F L. S S is th e fir s t lig h t s e p a r a te d fr o m th e o u tp u t o f th e p ic k u p le ns /s p a tia l filte r a s s e m b ly .

S S is s e p a r a te d u s ing a 48 8 nm dic h r o ic lo ng -p a s s ( 48 8 D L) filte r a t a 45-de g r e e a ng le to th e lig h t p a th ( s e e F ig u r e 5.3, F ig u r e 5.4, F ig u r e 5.6, a nd F ig u r e 5.5) . T h e 48 8 D L filte r r e fle c ts th e S S to th e S S s e ns o r b u t tr a ns m its flu o r e s c e nt lig h t o f lo ng e r w a v e le ng th s .

Fig ure 5.5 Sing le La ser Filter Block Config ura tion

b 48 8 D LP g 67 5 B P 1! 525 B P

c 50 0 LP h 7 10 D LP 1@ 530 S P

d 60 0 D LP i 7 55 LP 1# F lo w c e ll

e 645 D S P j 57 5 B P

f 620 B P 1) 550 D LP

b

cd

e f

g

h

i

j1)1!

1@1#

PN 624923B 5-7

OPERATION PRINCIPLESL IG H T COL L ECTION, SEP A R A TION A ND M EA SU R EM ENT 5

Fig ure 5.6 Dua l La ser Filter Block Config ura tion

Fluorescent Lig h t

T h e lig h t th e 48 8 D LP filte r tr a ns m its g o e s to a 50 0 nm lo ng p a s s filte r. T h is filte r b lo c k s a ny r e m a ining la s e r lig h t, tr a ns m itting o nly flu o r e s c e nt lig h t. T h e r e m a ining o p tic a l filte r s s e p a r a te th e lig h t fo r th e fiv e F L s e ns o r s . N ote: T h is flu o r e s c e nt lig h t c o lle c tio n de s c r ip tio n a p p lie s to th e D u a l La s e r F ilte r B lo c k Co nfig u r a tio n o nly .

T h e fir s t filte r in th e lig h t p a th ( 50 0 LP ) b lo c k s th e s ide s c a tte r lig h t fr o m th e A r g o n la s e r a nd tr a ns m its a ll th e flu o r e s c e nc e c o lo r s a nd th e s ide s c a tte r lig h t fr o m th e r e d la s e r. T h is a s s e m b ly s e p a r a te s th e c o lle c te d flu o r e s c e nc e s ig na l into fiv e c o lo r b a nds . P MT s 1 to 4 c o lle c t s ig na ls c e nte r e d a p p r o x im a te ly a t 525, 57 5, 610 a nd 67 5 nm . P MT 5 c o lle c ts a s ig na l c e nte r e d a p p r o x im a te ly a t 7 55 nm . Lo ng p a s s filte r s a r e u s e d to tr a ns m it th e s e c o lo r b a nds , b lo c k th e s ide s c a tte r lig h t fr o m th e o p tio na l r e d la s e r a nd p r o v ide a dditio na l b lo c k ing o f th e s ide s c a tte r lig h t fr o m th e A r g o n la s e r. T h e c o lo r b a nds a r e de s ig ne d to m e a s u r e flu o r e s c e nc e lig h t fr o m th e flu o r o c h r o m e s s u c h a s F I T C, P E , E CD , P C5 o r A P C, P C7 e x c ite d b y illu m ina tio n fr o m th e A r g o n a nd th e o p tio na l r e d la s e r s . D ic h r o ic filte r s a r e u s e d to s p lit c o lo r s a t 550 , 60 0 , 645 a nd 7 10 nm . P o s itio ns o f th e dic h r o ic filte r s h a v e b e e n e ffic ie ntly de s ig ne d to r e du c e th e nu m b e r o f o p tic a l s u r fa c e s flu o r e s c e nc e lig h t m u s t p a s s to r e a c h th e p h o to s e ns o r s . T h e ir lo c a tio ns r e la tiv e to th e o p tic a l a x is h a v e a ls o b e e n o p tim iz e d fo r lig h t to p a s s s y m m e tr ic a lly th r o u g h e a c h filte r. Y o u c a n indiv idu a lly inte rc h a ng e th e o p tic a l filte r s . A s m a ll p la te h o lds a ll th e o p tic a l filte r s u s e d to s e p a r a te flu o r e s c e nc e s ig na ls a llo w ing fo r e a s y inte rc h a ng e o f g r o u p s o f filte r s . T h e filte r s a r e p r e c is e ly m o u nte d in th e ir h o u s ing a nd th e filte r p la te is p r e c is e ly m o u nte d o n th e o p tic a l p la te . T h e r e is no ne e d to r e a lig n th e o p tic a l s y s te m w h e n th e filte r s o r th e filte r p la te a r e c h a ng e d.

b 48 8 D LP g 67 5 B P 1! 525 B P

c 50 0 LP h 7 10 D LP 1@ 530 S P

d 60 0 D LP i 7 55 LP 1# F lo w c e ll

e 615 D S P j 57 5 B P

f 620 S P 1) 550 D LP

b

cd

e f

g

h

i

j1)1!

1@1#

PN 624923B5-8

OPERATION PRINCIPLESSIG NA L P R OCESSING

5.6 SIGNAL PROCESSING

Volta g e Pulse Sig na ls

T h e Cy to m e te r h a s s e v e n s e ns o r s , e a c h g e ne r a ting a v o lta g e p u ls e s ig na l a s e a c h c e ll p a s s e s th r o u g h th e la s e r b e a m . A v o lta g e p u ls e s ig na l is p r o p o r tio na l to th e inte ns ity o f lig h t th e s e ns o r r e c e iv e d. T h e Cy to m e te r e le c tr o nic s a m p lifie s , c o nditio ns , inte g r a te s , a nd a na ly z e s th e s e p u ls e s .

Pea k Sig na l

F ig u r e 5.7 s h o w s h o w a p e a k v o lta g e p u ls e s ig na l fo r m s a s a c e ll c r o s s e s th e la s e r b e a m .

r P a r t I o f F ig u r e 5.7 s h o w s w h e n th e c e ll e nte r s th e la s e r b e a m a nd s o m e lig h t is s c a tte r e d.

r P a r t II o f F ig u r e 5.7 s h o w s w h e n th e c e ll is in th e c e nte r o f th e la s e r b e a m a nd th e s c a tte r e d lig h t, a nd th e r e fo r e , th e p u ls e h e ig h t, r e a c h e s a m a x im u m .

r P a r t II I o f F ig u r e 5.7 s h o w s w h e n th e c e ll le a v e s th e la s e r b e a m a nd th e s c a tte r e d lig h t de c r e a s e s .

T h e inte ns ity o f lig h t s c a tte r o r flu o r e s c e nc e de te r m ine s th e h e ig h t o f th e p e a k p u ls e ( s e e F ig u r e 5.7 ) . T h e tim e th e p a r tic le is in th e la s e r b e a m de te r m ine s th e w idth o f th e p u ls e . T h e r e fo r e , th e to ta l flu o r e s c e nc e ( inte ns ity a nd tim e ) de te r m ine s th e a r e a u nde r th e p u ls e . F ig u r e 5.8 s h o w s h o w th r e e c e lls w ith th e s a m e a m o u nt o f to ta l flu o r e s c e nc e b u t w ith diffe r e nt flu o r e s c e nc e inte ns itie s , p r o du c e diffe r e nt p e a k p u ls e s .

PN 624923B 5-9

OPERATION PRINCIPLESSIG NA L P R OCESSING 5

Fig ure 5.7 Volta g e Pulse Form a tion, Pea k Sig na l

b La s e r b e a m e T im e

c Ce ll f P u ls e h e ig h t

d V o lts

b c

d

e

b

d

e

c

b

c

d

e

f

I

II

III

PN 624923B5-1 0

OPERATION PRINCIPLESSIG NA L P R OCESSING

Integ ra l Sig na l

B e c a u s e th e to ta l flu o r e s c e nc e in a ll th r e e c e lls is th e s a m e , b u t th e dis tr ib u tio n is diffe r e nt, th e p u ls e c a n b e inte g r a te d to p r o du c e a n inte g r a l s ig na l ( s e e F ig u r e 5.8 ) .

T h e h e ig h t o f th e inte g r a l p u ls e is p r o p o r tio na l to th e to ta l flu o r e s c e nc e a nd is o b ta ine d w h e n th e c e ll e x its th e la s e r b e a m . T h e p u ls e h e ig h t, h o w e v e r, r e p r e s e nts th e m o s t inte ns e a m o u nt o f flu o r e s c e nc e p r o du c e d. T h e a r e a u nde r th e p u ls e is p r o p o r tio na l to th e to ta l flu o r e s c e nc e .

Fig ure 5.8 Integ ra l a nd Pea k Pulses

Am p lifica tion

S o m e v o lta g e p u ls e s m u s t b e a m p lifie d s o th a t th e c h a r a c te r is tic s o f th e c e lls c a n b e m e a s u r e d.

T h e s y s te m le ts y o u :

r Inc r e a s e th e g a in to line a r ly a m p lify th e inte g r a l a nd p e a k s ig na ls .

r Lo g a r ith m ic a lly tr a ns fo r m th e line a r da ta .

A lo g a r ith m ic tr a ns fo r m a tio n a c c e ntu a te s th e diffe r e nc e s b e tw e e n th e s m a lle r p u ls e s a nd r e du c e s th e diffe r e nc e s b e tw e e n th e la r g e r p u ls e s .

Sig na ls Genera ted

T h e ins tr u m e nt s e ns o r s c a n g e ne r a te th e s e s ig na ls ( inte g r a l u nle s s s ta te d o th e r w is e ; LO G s ta nds fo r lo g a r ith m ic ) :

b D ir e c tio n o f s a m p le flo w e V o lts h V o lts

c Ce ll in la s e r b e a m f T im e i T im e

d P e a k p u ls e s g Inte g r a l p u ls e s

b

c

d e

f

i

g h

PN 624923B 5-1 1

OPERATION PRINCIPLESP R OTOCOL S 5

r F S , F S LO G , F S P E A K

r S S , S S LO G , S S P E A K

r F L1, F L1 LO G , F L1 P E A K




r F L5, F L5 LO G , ( F L5 u s e s th e A U X s ig na l p a th w a y ) .

F o r a ny g iv e n s a m p le , th e ins tr u m e nt c a n c r e a te u p to e ig h t o f th e s e s ig na ls , p lu s th e P r is m , T IME , a nd R A T I O p a r a m e te r s .

5.7 PROTOCOLSA P r o to c o l is a c o lle c tio n o f info r m a tio n a b o u t h o w th e Cy to m e te r a nd CX P s o ftw a r e is s e t u p . P r o to c o ls c a n b e c r e a te d fo r a c q u ir ing s a m p le s o r a na ly z ing s to r e d da ta .

It c o nta ins th e fo llo w ing info r m a tio n:

r D is p la y c o nfig u r a tio n

r P a r a m e te r na m e s a nd c o nfig u r a tio n

r R e g io ns

r G a te s

r Co lo r de finitio ns

r S ta tis tic s

r Ins tr u m e nt s e tting s

r R e p o r ting T e m p la te s

Sp ecia l Protocols a nd Pa nels

T h e r e a r e th r e e c a te g o r ie s o f s p e c ia l P r o to c o ls : Q C, Cle a ns e a nd A S .

Q C P r o to c o ls ( E x p o r t) : A ny P r o to c o l na m e b e g inning w ith th e c h a r a c te r s “ Q C” ( u p p e r- c a s e o nly ) o v e r-r ide s th e c u r r e nt E x p o r t D a ta F o r m a t s e tting o n th e W o r k s p a c e P r e fe r e nc e s / P u b lis h ta b c a u s ing P u b lis h e d da ta to b e fo r m a tte d a s a s ing le r o w o f v a lu e s . In th e c a s e th a t th e P u b lis h D a ta to MS E x c e l o p tio n is s e le c te d, th is p r o g r e s s iv e ly b u ilds a ta b le o n a s ing le s p r e a ds h e e t.

Cle a ns e P r o to c o ls : A ny P r o to c o l na m e b e g inning w ith th e w o rd “ Cle a ns e ” o v e r r ide s th e O u tp u t O p tio ns s e tting fo r S a v e LMD o n th e W o r k s p a c e P r e fe r e nc e s - A c q u is itio n O p tio ns ta b . T h is a llo w s y o u to c r e a te p r o to c o ls th a t do no t w r ite a lis tm o de file to dis k o n c o m p le tio n.

A S P r o to c o ls : T h e r e is a s e r ie s o f s p e c ia l p r o to c o ls w h ic h inc lu de “ A S ” in th e ir na m e s . T h e s e a r e c o m p o ne nts o f th e A u to S e tu p a p p lic a tio ns w h ic h o p e r a te w ith th e A u to S e tu p W iz a rd to s e t u p V o lts , G a ins a nd Co m p e ns a tio n.

Sp ecia l Pa nels

T h e c le a ns e p a ne l u tiliz e s tw o c le a ns e p r o to c o ls ( ‘c le a ns e b le a c h .P R O ’ a nd ‘c le a ns e di w a te r.P R O ) th a t y o u s h o u ld u s e a t th e e nd o f th e w o r k ing da y o r s h ift o n y o u r F C 50 0 .

PN 624923B5-1 2

OPERATION PRINCIPLESA U TOM A TED SOFTW A R E F EA TU R ES

5.8 AUTOMATED SOFTWARE FEATURESCX P s o ftw a r e c o nta ins th e fo llo w ing a u to m a te d s o ftw a r e fe a tu r e s .

r A u to S e tu p W iz a rd

r Q u ic k CO MP

r Q u ic k S E T

r A u to m a tic G a te Cre a tio n

r A u to m a tic G a te Ma inte na nc e

r E llip tic a l a nd Co nto u r A u to G a ting

r A u to MA T O R A na ly s is

5.9 PARAMETERS

AUX ilia ry Pa ra m eter

T h e A U X s ig na l is a v a ila b le to c o lle c t a P e a k s ig na l fr o m a ny o f th e de te c te d p a r a m e te r s w h e n a na ly z ing u p to fo u r c o lo r s . W h e n u s ing F L5 o r a na ly z ing fiv e c o lo r s th e A U X s ig na l is no t a v a ila b le . F o r a ny g iv e n s a m p le , o nly o ne o f th e fo llo w ing s ig na ls c a n b e a s s ig ne d to A U X :

r F S , F S P E A K

r S S , S S P E A K

r F L1, F L1 P E A K




Wh en to Use th e AUX Pa ra m eter

W h e n y o u s p e c ify b o th line a r a nd lo g a r ith m ic a m p lific a tio n o f th e s a m e s ig na l, th e g a in fo r th e line a r a m p lific a tio n is a ls o a p p lie d to th e lo g a r ith m ic a m p lific a tio n. Ho w e v e r, w h e n y o u s p e c ify o nly lo g a r ith m ic a m p lific a tio n o f a s ig na l, th e ins tr u m e nt a u to m a tic a lly s e ts th e g a in fo r th a t s ig na l to 1.0 , a nd y o u c a nno t c h a ng e it.

U s e th e A U X p a r a m e te r w h e n y o u w a nt to :

r A m p lify a s ig na l a t tw o diffe r e nt g a ins .

r O b s e r v e a p e a k s ig na l o r inte g r a l s ig na l a t th e s a m e tim e a s th e lo g s ig na l.

r O b s e r v e a p e a k s ig na l a t th e s a m e tim e a s th e inte g r a l s ig na l.

A ls o , th e A U X p a r a m e te r m a y b e u s e d fo r do u b le t dis c r im ina tio n. A s s ig n a p e a k flu o r e s c e nc e s ig na l to A U X s o y o u c a n m e a s u r e p e a k v s . inte g r a l flu o r e s c e nc e .

TIME Pa ra m eter

T h e T IME p a r a m e te r is th e a m o u nt o f tim e , in s e c o nds , th e ins tr u m e nt a c q u ir e s da ta . I t is dis p la y e d o n th e p lo t a x is in 1-s e c o nd r e s o lu tio n. T h e a x is la b e ls v a r y, de p e nding o n p lo t r e s o lu tio n a nd s to p tim e ( du r a tio n) .

PN 624923B 5-1 3

OPERATION PRINCIPLESP L OT D ISP L A Y 5

T h e m inim u m s to p tim e is 10 s e c o nds , th e m a x im u m s to p tim e ( m a x im u m du r a tio n) is 99,999 s e c o nds a nd th e de fa u lt s to p tim e is 30 0 s e c o nds ( 5 m inu te s ) .

W h e n y o u a s s ig n th e T IME p a r a m e te r to a p lo t a x is , th e div is io ns o n th e a x is c h a ng e a c c o rding ly .

T o find th e tim e ( in s e c o nds ) p e r c h a nne l in a o ne -p a r a m e te r h is to g r a m , div ide th e s to p tim e ( in s e c o nds ) b y 1,0 24 ( 0 .0 0 1 s e c o nd = 1.0 m s ) .

F o r a tw o - p a r a m e te r p lo t, div ide b y 64, 128 , 256 o r 512 de p e nding u p o n th e p lo t r e s o lu tio n y o u a r e u s ing .

RATIO Pa ra m eter

T h e R A T I O p a r a m e te r is c a lc u la te d, no t a c q u ir e d dir e c tly . W h e n y o u s e le c t a p a r a m e te r, y o u s p e c ify w h ic h s ig na l is th e nu m e r a to r a nd w h ic h is th e de no m ina to r .

A r a tio o f 1 is a t c h a nne l 1,0 23. If y o u a s s ig n R A T IO to a p lo t a x is , R A T IO e v e nts a p p e a r a t a lo w e r c h a nne l if th e inte ns ity o f th e nu m e r a to r s ig na l is le s s th a n th e de no m ina to r s ig na l.

T o c a lc u la te th e a c tu a l r a tio a t a p a r tic u la r inte ns ity fo r a o ne -p a r a m e te r h is to g r a m , div ide th e inte ns ity b y 1,0 24. F o r a tw o - p a r a m e te r p lo t, div ide b y 64, 128 , 256 o r 512 de p e nding u p o n th e p lo t r e s o lu tio n y o u a r e u s ing .

5.10 PLOT DISPLAYT h e r e s u lts o f s a m p le a na ly s is a p p e a r o n th e W o r k s ta tio n s c r e e n a s g r a p h s c a lle d p lo ts . Y o u a s s ig n th e p a r a m e te r s to th e p lo t a x e s . P lo ts c a n b e dis p la y e d in b la c k a nd w h ite o r c o lo r a s :

r S ing le - p a r a m e te r h is to g r a m p lo t

r D u a l- p a r a m e te r do t p lo t

r D u a l- p a r a m e te r c o nto u r p lo t ( CX P A na ly s is S o ftw a r e o nly )

r D u a l- p a r a m e te r de ns ity p lo t

r D u a l- p a r a m e te r s u r fa c e p lo t

r S ing le - p a r a m e te r P r is m p lo t

r O v e r la y e d h is to g r a m p lo ts ( CX P A na ly s is S o ftw a r e o nly )

r 3-D to m o g r a m p lo t ( CX P A na ly s is S o ftw a r e o nly ) .

D u a l- p a r a m e te r p lo ts c a n b e dis p la y e d in 64 x 64, 128 x 128 , 256 x 256, o r 512 x 512 r e s o lu tio n. P lo ts c a n b e dis p la y e d o n a b la c k o r w h ite b a c k g r o u nd.

PRISM

P r is m is u s e d to a na ly z e m u ltic o lo r im m u no flu o r e s c e nc e s a m p le s . W ith m u ltic o lo r im m u no flu o r e s c e nc e a c e ll is e ith e r p o s itiv e o r ne g a tiv e fo r e a c h o f tw o , th r e e , fo u r o r fiv e c e ll s u r fa c e m a r k e r s . A p a r tic u la r c o m b ina tio n is c a lle d a p h e no ty p e . P r is m a llo w s y o u to dis p la y p e rc e nta g e s o n a ll p h e no ty p ic p o p u la tio ns in a s ing le p lo t. It is s o ftw a r e de r iv e d a nd c a n b e a c q u ir e d in e ith e r r u n tim e o r lis tm o de .

RATIONumerator

Denominator----------------------------------- 1024×=

PN 624923B5-1 4

OPERATION PRINCIPLESP L OT D ISP L A Y

P r is m is a v a ila b le o n u p to fiv e p a r a m e te r s . T IME , R A T IO , a nd P r is m its e lf c a nno t b e u s e d fo r P r is m . A ll o th e r s ig na ls c a n b e u s e d fo r P r is m . G e ne r a lly , F L1, F L2, F L3, F L4, a nd F L5 a r e u s e d.

A P r is m p lo t s h o w s a s p ik e o r p o p u la tio n fo r e a c h a ntib o dy c o m b ina tio n w ith a p e r c e nt o f th e to ta l th a t r e p r e s e nts th e p e rc e nt o f th e to ta l e v e nts in th e P r is m p lo t. S e e F ig u r e 5.9.

Fig ure 5.9 Prism Plot

S e e P r is m P lo t fo r ins tr u c tio ns o n u s ing P r is m .

PN 624923B 5-1 5


Reg ions

T o a na ly z e da ta o r g a te p lo ts , y o u m u s t fir s t c r e a te a nd a s s ig n r e g io ns to th e s e ta s k s . Y o u c a n c r e a te fiv e diffe r e nt ty p e s o f r e g io ns ( s e e R e g io ns Intr o du c tio n) . T h e r e g io n ty p e s a r e :

r Line a r

r R e c ta ng u la r

r Q u a dr a nt

r P o ly g o na l

r E llip tic a l

r Co nto u r

r O n O v e r la y e d h is to g r a m s o nly

r Link e d m a r k e r

r N o n-link e d m a r k e r

O nc e a r e g io n is c r e a te d it c a n b e a s s ig ne d to fu nc tio n in a s p e c ific w a y . T h e fu nc tio ns th a t a r e a v a ila b le inc lu de :

r A na ly s is

r P r im e

r CA L ( c a lib r a tio n)

r G a ting

r Lis tm o de g a ting ( LIV E G A T E )

r Co nto u r A u to G a te

r E llip tic a l A u to G a te

r P o s itiv e s a na ly s is

r Minim u m Co u nt.

T h e s e fu nc tio ns a r e no t a v a ila b le fo r a ll r e g io n ty p e s .

Ga ting

T h e s o ftw a r e le ts y o u u s e g a ting to s p e c ify th a t o nly c e r ta in c e lls a r e to b e a na ly z e d. A g a te c a n b e de fine d a s th e c e lls th a t a r e ins ide o r o u t o f o ne o r m o r e r e g io ns .

Da ta Stora g e

S a m p le r e s u lts c a n b e p r inte d o u t, s a v e d to a dis k e tte o r o th e r r e m o v a b le m e dia , s a v e d to a lo c a l h a rd dr iv e o r s a v e d to a ne tw o r k dr iv e . Y o u c a n s to r e s a m p le r e s u lts in th e fo r m o f a lis t o f th e m e a s u r e m e nts fr o m e a c h c e ll, c a lle d lis tm o de da ta . Lis tm o de da ta c a n b e r e p la y e d into p lo ts o r a rc h iv e d fo r a na ly s is la te r. P lo ts c a n a ls o b e s a v e d to a file .

PN 624923B5-1 6


Histog ra m Sta tistics

Linea r Reg ion Sta tistics

F o r line a r s ig na ls , s ta tis tic s fo r h is to g r a m r e g io ns a r e c a lc u la te d a s fo llo w s :

N u m b e r = N u m b e r o f c e lls in th e r e g io n

Mo de = Inte ns ity c o nta ining th e la r g e s t nu m b e r o f c e lls w ith in th e r e g io n.

W h e r e δ = S ta nda rd de v ia tio nCV = Co e ffic ie nt o f V a r ia tio nMH = Hig h r e g io n m a r k e r c h a nne l

ML = Lo w r e g io n m a r k e r c h a nne l

Cn = R a w dig ita l c h a nne l nu m b e r

Cc h = Ch a nne l c o u nt

N = Inte g r a l o f a ll c o u nts b e tw e e n & inc lu ding m a r k e r c h a nne ls

= Me a n c h a nne l nu m b e r ( r o u nde d to ne a r e s t w h o le c h a nne l)

F o r Me a n a nd S D , th e s u m m a tio ns a r e p e r fo r m e d o v e r a ll th e c h a nne ls th a t lie w ith in th e r e g io n.

Me dia n inte ns ity = th e s m a lle s t inte ns ity s u c h th a t s u m ( inte ns ity ) ≥

S u m ( inte ns ity ) = s u m o f e v e nts fr o m th e lo w e r e dg e o f th e r e g io n to th e inte ns ity

totalNumb er of ev ents in reg ion

Total numb er of ev ents in th e g ated d is p lay-------------------------------------------------------------------------------------------------------- 100×=

totalNumb er of c ells in reg ion

Total numb er of c ells in th e file---------------------------------------------------------------------------- 100×=

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PN 624923B 5-1 7


W h e r e F W HM= th e F u ll W idth Ha lf Ma x v a lu e o f a N o r m a l o r G a u s s ia n p e a k .

Log Reg ion Sta tistics

T h e m e th o d o f g e o m e tr ic c a lc u la tio ns o f Me a ns de p e nds o n y o u r A dv a nc e d S ta tis tic s Co nfig u r a tio n.

Log -Log Mea n Meth od

W e ig h ting e a c h c h a nne l a c c o rding to its fa c e v a lu e a s g iv e n b y th e lo g s c a le p e r fo r m s th is c a lc u la tio n o f Me a n.

E x a m p le

W h e n u s ing a fo u r-de c a de u nc a lib r a te d lo g a r ith m ic dis p la y r e p o r ting o u tp u t a s Ch a nne ls ,

y o u m ig h t g iv e c h a nne l z e r o ( th e lo w e s t c h a nne l) a v a lu e 10 -1 = 0 .1 a nd o n a 10 24 c h a nne l

dis p la y , c h a nne l 10 23 is g iv e n th e v a lu e 10 -3 = 10 0 0 . A ll r a w dig ita l c h a nne l v a lu e s a r e c o nv e r te d to a n a b s o lu te v a lu e ( R e la tiv e Ch a nne l N u m b e r ) in th e r a ng e 0 .1 to 10 0 0 b e fo r e c a lc u la tio ns a r e p e r fo r m e d fo r th e m e a n.

T h e m e a n c a n th e n b e r e p o r te d in Ch a nne l nu m b e r s in th e r a ng e 0 to 10 23 th u s :

W h e r e :Cn = R a w dig ita l c h a nne l nu m b e r


Cd = Ch a nne ls p e r de c a de



F o r Ca lib r a te d v a lu e s , th e c a lc u la tio n is r e p o r te d a s th e c a lib r a te d v a lu e in th e r a ng e Lo g O ffs e t to

T h e lo g o ffs e t c a nno t b e z e r o s inc e a lo g o ffs e t o f z e r o is 10 ( m inu s infinity ) , w h ic h is no t p o s s ib le to r e p r e s e nt o n p a p e r o r o n s c r e e n. CX P s o ftw a r e do e s no t a llo w lo g o ffs e t v a lu e s o f le s s th a n 0 .1. N u m b e r s e nte r e d in th e lo g a r ith m ic o ffs e t a r e c la m p e d a t 0 .1 if a nu m b e r le s s th a n th is is e nte r e d.

H alf P eak C VF W H M

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PN 624923B5-1 8


Lin-Log Mea n Meth od

T h is m e th o d o f Me a n c a lc u la tio n is w o r k e d o u t b y p e r fo r m ing a s im p le a r ith m e tic m e a n c h a nne l c a lc u la tio n o n r a w da ta a c c o rding to its c h a nne l v a lu e in th e r a ng e 0 to 10 23 ( fo r 10 24 c h a nne l da ta ) . T h e Me a n v a lu e c a n th e n b e r e p o r te d a s c h a nne ls ( in th e r a ng e 0 to 10 23) o r c o nv e r te d to th e R e la tiv e Ch a nne l v a lu e .

U s ing th e s a m e da ta a s in th e p r e v io u s e x a m p le …

Lin-Lo g Me a n m e th o d r e p o r te d in Ch a nne l nu m b e r s in th e r a ng e 0 -10 23 th u s :

W h e r e :Cn = R a w dig ita l c h a nne l nu m b e r


Cd = Ch a nne ls p e r de c a de



F o r c a lib r a te d v a lu e s , th e Me a n c a lc u la tio n is r e p o r te d a s a v a lu e in th e r a ng e Lo g O ffs e t to

( Lo g o ffs e t* 10 D e c a de s F u ll S c a le ) th u s :

T h e lo g o ffs e t c a nno t b e z e r o s inc e a lo g o ffs e t o f z e r o is 10 ( m inu s infinity ) . S inc e th e g r a p h ic a l s c a le is a c tu a lly p r o p o r tio na l to th e e x p o ne nt, it is no t p o s s ib le to r e p r e s e nt th is o n p a p e r o r o n s c r e e n. B e c a u s e o f th is p r o b le m , CX P s o ftw a r e do e s no t a llo w lo g o ffs e t v a lu e s o f le s s th a n 0 .1. N u m b e r s e nte r e d in th e Lo g a r ith m ic O ffs e t e dit w indo w a r e c la m p e d a t 0 .1 if a nu m b e r le s s th a n 0 .1 is e nte r e d.

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PN 624923B 6-1

66TROUBLESHOOTING

6.1 PRECAUTIONS/HAZ ARDS

La ser/Ra d ia tion Preca utions

T h e Cy to m e te r a nd th e MCL b a r-c o de r e a de r e a c h c o nta in a la s e r. T h e Cy to m e te r c a n a ls o inc lu de a n o p tio na l s e c o nd la s e r. B e c k m a n Co u lte r 's de s ig n a nd m a nu fa c tu r e o f th e ins tr u m e nt c o m p lie s w ith th e r e q u ir e m e nts g o v e rning th e u s e a nd a p p lic a tio n o f a la s e r a s s p e c ifie d in r e g u la to r y do c u m e nts is s u e d b y th e :

r U .S . D e p a r tm e nt o f He a lth a nd Hu m a n S e r v ic e s a nd

r Ce nte r fo r D e v ic e s a nd R a dio lo g ic a l He a lth ( CD R H) .

In c o m p lia nc e w ith th e s e r e g u la to r y do c u m e nts , e v e r y m e a s u r e h a s b e e n ta k e n to e ns u r e th e h e a lth a nd s a fe ty o f u s e r s a nd la b o r a to r y p e r s o nne l fr o m th e p o s s ib le da ng e r s o f la s e r u s e .

U s e th e ins tr u m e nt a c c o rding to th e info r m a tio n in th e m a nu a ls .

U s e o f c o ntr o ls o r a dju s tm e nts o r p e r fo r m a nc e o f p r o c e du r e s o th e r th a n th o s e s p e c ifie d h e r e in m ig h t r e s u lt in h a z a rdo u s r a dia tio n e x p o s u r e .

T o e ns u r e y o u r s a fe ty , th e Cy to m e te r la s e r s a r e c o v e r e d w ith p r o te c tiv e s h ie lds . D o no t r e m o v e th e s e s h ie lds .

N o u s e r- s e r v ic e a b le a s s e m b lie s a r e a c c e s s ib le . D o no t a tte m p t to r e m o v e th e la s e r o r o p e n it.

T h e ins tr u m e nt h a s c o m p o ne nts th a t a r e da ng e r o u s to th e o p e r a to r. If a ny a tte m p t h a s b e e n m a de to de fe a t a s a fe ty fe a tu r e , o r if th e ins tr u m e nt fa ils to p e r fo r m a s de s c r ib e d in its m a nu a ls , dis c o nne c t th e p o w e r a nd c a ll y o u r B e c k m a n Co u lte r R e p r e s e nta tiv e .

La ser Wa rning La b els

CD R H-r e q u ir e d w a rning la b e ls a r e p la c e d ne a r o r o n c o v e r s th a t, if r e m o v e d, m ig h t e x p o s e la s e r r a dia tio n. T h e y a r e a ls o p la c e d ne a r o p e ning s th a t, if lo o k e d into , m ig h t e x p o s e y o u to la s e r r a dia tio n.

CD R H-r e q u ir e d w a rning la b e ls a r e lo c a te d:

S e e F ig u r e 6.1 fo r th e S e ns ing Co m p a r tm e nt c o v e r w a rning la b e l.

S e e F ig u r e 6.2 fo r th e O p tic a l A r e a ( fr o nt v ie w ) w a rning la b e ls .

S e e F ig u r e 6.3 fo r th e O p tic a l A r e a ( inte r io r v ie w ) w a rning la b e ls .

S e e F ig u r e 6.4 fo r th e A r g o n La s e r He a d w a rning la b e ls .

S e e F ig u r e 6.5 fo r th e o p tio na l r e d S o lid-s ta te La s e r w a rning la b e ls .

S e e F ig u r e 6.6 fo r th e lo c a tio n o f th e o p tio na l r e d S o lid-s ta te La s e r in th e ins tr u m e nt.

S e e F ig u r e 6.7 fo r th e o p tio na l r e d He liu m -N e o n La s e r w a rning la b e ls .

S e e F ig u r e 6.8 fo r th e lo c a tio n o f th e o p tio na l r e d He liu m -N e o n La s e r in th e ins tr u m e nt.

S e e F ig u r e 6.9 fo r th e Cy to m e te r b a c k p a ne l la s e r la b e ls .

S e e F ig u r e 6.10 fo r th e MCL p r o b e h o u s ing w a rning la b e ls .

S e e F ig u r e 6.11 fo r th e MCL b a r- c o de r e a de r w a rning la b e ls .

PN 624923B6-2

TROUBLESHOOTINGP R ECA U TIONS/H A Z A R D S

Fig ure 6.1 La ser La b els on th e Sensing Com p a rtm ent Cov er

7 27 2009A

AVOID EYE OR SKINEXPOSURE TO DIRECT ORSCATTERED RADIATION

VISIB L E AND/OR INVISIB L EL ASER RADIATION W H EN OPENL ASER RADIATION

AVOID EXPOSURE TO B EAMCL ASS 3 B L ASER PRODUCT

PN 624923B 6-3

TROUBLESHOOTINGP R ECA U TIONS/H A Z A R D S 6

Fig ure 6.2 La ser La b els in th e Op tica l Area , Front View

A V OID EX POSU RE

A V OID EX POSU REVISIB L E AND/ORINVISIB L E L ASER

RADIATION ISEM ITTED F ROM

TH IS APERTURE.

VISIB L E AND/OR INVISIB L EL ASER RADIATION W H EN OPEN


7 2 7 2 0 0 6 A

PN 624923B6-4


Fig ure 6.3 La ser La b els in th e Op tica l Area , Interior View

7 27 407 8A


VISIB L E AND/OR INVISIB L EL ASER RADIATION W H EN OPEN

CA U TIONLa s e r r a d ia tio n w h e n o p e n

a n d in te r lo c k d e fe a te d .

D O N OT STA RE IN TO B EA M.

A V OID EX POSU RE

VISIB L E AND/OR INVISIB L EL ASER RADIATION IS EM ITTED

F ROM TH IS APERTURE.

PN 624923B 6-5


Fig ure 6.4 La ser La b els on th e Arg on La ser Hea d

Fig ure 6.5 La ser La b els for Op tiona l Red Solid -sta te La ser

7 27 2007 A

L ASER RADIATION-AVOIDDIRECT EXPOSURE TO B EAM

ARG ON/.5 0 W ATT C.W .

CL ASS IIIb L ASER PRODUCTA V OID EX POSU RE

L ASER RADIATIONIS EM ITTED F ROMTH IS APERTURE

L ASER RADIATION W H EN OPEN

AVOID DIRECT EXPOSURE TO B EAM

Redflame

CLO

SE

A V OID EX P OSU R E

LA SER R A D IA T ION EM IT T ED

F R OM T H IS A P ER T U R E

LASER RADIATION

W A V ELEN G T H /P OW ER

P A R T N U M B ER /R EV ISION

SER IA L N U M B ER

B EA M

OP T ION S

A V OID EX P OSU R E T O B EA M CLA SS lllB

CD R H LA SER P R OD U CT CLA SS 3 b

LA SER P ER EN 6 0 8 2 5 -1

P EA K P OW ER 3 0 m W

2 3 0 3 LIN D B ER G H ST .

A U B U R N , CA 9 5 6 0 2 -9 5 9 5

T H IS P R OD U CT D OES N OT COM F OR M T O

T H E A P P LICA B LE ST A N D A R D S OF 2 1 CF R 1 0 4 0 .

COH ER EN T

1 0 0 8 0 5 5 3 /A B

6 3 8 n m /2 2 m W

X X X X X

1 m m CIR CU LA R

N /A

7 3 1 8 2 1 0 B

PN 624923B6-6


Fig ure 6.6 Loca tion of th e Op tiona l Red Solid -sta te La ser in th e Instrum ent, Rea r View

Fig ure 6.7 La ser La b els for Op tiona l Red Helium -Neon La ser

b S olid -s tate las er h ead c M ounting p late d Protective h ous ing

FI BE

RAN

RE

SC OP E

SCOPE

FLOWCEL WAST

WAST PRESSURE VAC UUM

LOGIC ANALOG OPTI

M anu fa ct ur ed

D A T E b y CO U L T E R CO R P O R AT IO

1 1 800 S . W . 1 4 7 AV E

M I AM I, FL 33 1 96- 250

T H IS L AS E R PR O D U C T CO NF O R M S T O T H E PR O V IS IO NS O F 21 CF R .

S U BC H A PT E R J, S E CT IO NS 1 0 40 .1 AN D 040. 1 1 .

POWER MO DUL CONTRO

VI SI B L E AN D/ OR IN VI SI B L

L A SER RA DI AT IO N W H EN OP EN

MCL PO WE

5 5 7 1 0 1 9 A

COHERENT

cb d

JDS Uniphase1 096 M ellon AvenueM anteca, CA 95337

M O D E L M ANU FACT U R E D S E R IAL NO .

T H IS L AS E R PR O D U CT CO M PL IE SW IT H 21 CFR 1 040 AND

IE C 825-1 : 1 993.

S E E INS T AL L AT IO N INS T R U CT IO NS .

PAT E NT NO SM AD E

INU S A

L AS E R R AD IT IO NAV O ID E X PO S U R E T O BE AM

CL AS S 3B L AS E R PR O D U CT (IE C)CL AS S IIIb L AS E R PR O D U CT (CD R H )

633 nm / 35 mW

LAS

ER R

AD

ATIO

NIS

EM

ITTE

D F

RO

MTH

IS A

PER

TUR

E

AV

OID

EX

PO

SU

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CLOSED OPENBEAM SHUTTER

PL ANE O F PO L AR IZ AT IO N

JDS Unip

hase

7 27 2002A

PN 624923B 6-7


Fig ure 6.8 Loca tion of th e Op tiona l Red Helium -Neon La ser in th e Instrum ent, Rea r View

Fig ure 6.9 La ser La b els on th e Cy tom eter Ba ck Pa nel

b H eNe las er h ead c M ounting p late d Protective h ous ing

FI BE

RAN

RE

SC OP E

SCOPE

FLOWCEL WAST


LOGIC ANALOG OPTI

M anu fa ct ur ed

D A T E b y

CO U L T E R CO R P O R AT IO

1 1 800 S . W . 1 4 7 AV E

M I AM I, FL 33 1 96- 250


S U BC H A PT E R J, S E CT IO NS 1 0 40 .1

AN D 040. 1 1 .

POWER MO DUL CONTRO



MCL PO WE

7 2 7 2 0 0 8 Acb d

M a n u fa c tu re d :DATE b y :COUL TER CORPORATION1 1 8 0 0 S.W . 1 4 7 AVEM IAM I, F L 3 3 1 9 6 -2 5 0 0

TH IS L ASER PRODUCT CONF ORM STO TH E PROVISIONS OF 2 1 CF R.SUB CH APTER J , SECTIONS 1 0 4 0 .1 0AND 1 0 4 0 .1 1 .

VISIB L E AND/OR INVISIB L E

L ASER RADIATION W H EN OPEN

AVOID EYE OR SKIN

EXPOSURE TO DIRECT OR

SCATTERED RADIATION

FI BE

RAN

RE

SC OP E

SCOPE

FLOWCEL

WAST


LOGIC ANALOG OPTI

M anu fa ct ur ed

D A T E b y CO U L T E R CO R P O R AT IO

1 1 800 S . W . 1 4 7 AV E

M I AM I, FL 33 1 96- 250


S U BC H A PT E R J, S E CT IO NS 1 0 40 .1 AN D 040. 1 1 .

POWER MO DUL

CONTRO



AV OI D EY E OR SK IN

EX POS URE TO DI RE CT OR

SC AT TE RE D RA DI AT IO

MCL PO WE

b

c

b L ab el is ap p lied only w h en an op tional s econd las er is ins talled .

c T h is cover is only p res ent w h en ins trument contains an op tional H eNe las er.

PN 624923B6-8


Fig ure 6.10 La ser La b els on th e MCL Prob e Housing Cov er

CA U TIONL ASER L IG H T DO NOT STARE INTO B EAM

6 7 0 n m DIODE L ASER1 .0 M IL L IW ATT M AXIM UM CL ASS II L ASER PRODUCT

CA U TIONLa s e r r a d ia tio n w h e n o p e n

a n d in te r lo c k d e fe a te d .

D O N OT STA RE IN TO B EA M.

CLA SS 1 LA SER PROD U CT

A V OID EX POSU RE

VISIB L E AND/OR INVISIB L EL ASER RADIATION IS EM ITTED

F ROM TH IS APERTURE.

5 5 7 0 0 0 2 B

PN 624923B 6-9


Fig ure 6.11 La ser La b els on th e MCL Ba r-Cod e Rea d er

7 27 2001 A

AVOID EX POSURE LASER LIGHT IS EMITTEDFROM THIS APERTURE

CAUTION LASER LIGHT WHEN OPEN-DO NOT STARE INTO BEAM.PRODUCT CONFORMS TO USA DHHS 21 CFR SUBCHAPTER "J "

CAUTION VORSICHTL AS E R L IG H T D O NO T S T AR E INT O BE AM

67 0 nm L AS E R D IO D E1 .0 M IL L IW AT T M AX

CL AS S II L AS E R PR O D U CT

AVOID EXPOSURE

LASER LIGHT IS

EMIT

TED

FROM THIS

APERTURE

CAUTION LASER LIG

HT WHEN O

PEN-DO N

OT STARE IN

TO BEAM

.

PRODUCT CONFORM

S TO USA D

HHS 21 CFR S

UBCHAPTER "J"

CAUTION

VORSICHT

SER

IAL

NU

MBE

R/M

OD

EL N

UM

BER

MAN

UFA

CTU

RED

MIC

RO

SCAN

FIS

CG

D/R

DR

DY

PWR

US

UL

SERIAL NUMBER/MODEL NUMBER

MANUFACTURED

MICROSCAN

FIS

CGD/RD RDY PWR

USUL

PN 624923B6-1 0


Wa rning La b els on UPS

T h e la b e l lo c a te d ne a r th e c irc u it b r e a k e r s o n th e U niv e r s a l P o w e r S u p p ly ins tr u c ts th e u s e r to r e fe r to p r o du c t do c u m e nta tio n b e fo r e r e s e tting th e c irc u it b r e a k e r s .

T h e la b e l lo c a te d ne x t to th e p o w e r s w itc h o n th e U niv e r s a l P o w e r S u p p ly ins tr u c ts th e u s e r to r e fe r to p r o du c t do c u m e nta tio n b e fo r e p o w e r ing u p th e ins tr u m e nt.

Fig ure 6.12 Interna tiona l Wa rning Sy m b ol Loca tions

SYSTEM POWER

MCL POWER

WATER TRAP

VAC TRAP

AIR FILTER

V A C

F ILTER

SY S VAC

SY S PRESS

30 PSI

PRESSADJ

SYSTEM POWER

MCL POWER

WATERTRAP

VAC TRAP

AIR FILTER

V A C

F ILTER

SY S VAC

SY S PRESS

30PSI

PRESSADJ

PN 624923B 6-1 1

TROUBLESHOOTINGM A INTENA NCE SCH ED U L ES 6

Disp osa l Preca ution

6.2 MAINTENANCE SCHEDULES

Clea ning Sch ed ule

S e e T a b le 6.1 fo r th e c le a ning s c h e du le .

Rep la cem ent Sch ed ule

T h e s h e a th flu id filte r ne e ds to b e r e p la c e d e v e r y 6 m o nth s . A ll o th e r r e p la c e m e nt a nd a dju s tm e nt p r o c e du r e s s h o u ld b e do ne o n a n a s ne e de d b a s is .

6.3 ERROR MESSAGESS e e T a b le 6.2 fo r a lis t o f a ll o f th e e r r o r m e s s a g e s .

Disp la y Loca tions

E r r o r m e s s a g e s c a n a p p e a r a t th e b o tto m o f th e CX P s o ftw a r e s c r e e n, in th e Cy to m e te r S ta tu s Me s s a g e s s c r e e n, a nd in th e c y to m e te r.lo g file .

CX P Softw a re Screen

A t th e b o tto m o f th e CX P s o ftw a r e s c r e e n, th e la s t e r r o r m e s s a g e a p p e a r s . S e e F ig u r e 6.13.

N ote: I f m u ltip le e r r o r m e s s a g e s a r e p o s te d a t th e s a m e tim e , o nly th e la s t o ne p o s te d a p p e a r s h e r e . A ll o f th e e r r o r m e s s a g e s p o s te d a p p e a r in th e Cy to m e te r S ta tu s Me s s a g e s s c r e e n a nd th e c y to m e te r.lo g file .

WARNING R is k of b ioh az ard ous contamination if y ou h ave s kin contact w ith th e w as te container, its contents , and its as s ociated tub ing . T h e w as te container and its as s ociated tub ing mig h t contain res id ual b iolog ical material and mus t b e h and led w ith care. Clean up s p ills immed iately . D is p os e of th e contents of th e w as te container in accord ance w ith y our local reg ulations and accep tab le lab oratory p roced ures

Ta b le 6.1 Clea ning Sch ed ule

Com p onent Da ily Week ly Month ly Ev ery 60 Da y s As Need ed

CL E AN T H E AIR FIL T E R S - c - - -

CL E AN T H E CL E ANING AG E NT CO NT AINE R

- - - c -

CL E AN T H E M CL S AM PL E H E AD AND T H E S AM PL E PR O BE

c c - - -

CL E AN T H E S AM PL ING S Y S T E M c - - - -

CL E AN T H E S H E AT H FL U ID CO NT AINE R

- - c - -

CL E AN T H E V ACU U M T R AP - - - - c

- = Not Ap p licab le

PN 624923B6-1 2

TROUBLESHOOTINGER R OR M ESSA G ES

Fig ure 6.13 Error Messa g e on CX P Softw a re Screen, Ex a m p le

Cy tom eter Sta tus Messa g es Screen

In th e Cy to m e te r S ta tu s Me s s a g e s s c r e e n, th e la s t e r r o r m e s s a g e a nd a ll u nc le a r e d e r r o r m e s s a g e s a p p e a r. S e e F ig u r e 6.14.

r T o a c c e s s th e Cy to m e te r S ta tu s Me s s a g e s s c r e e n, do u b le c lic k th e e r r o r m e s s a g e a t th e b o tto m o f th e CX P s o ftw a r e s c r e e n.

r Clea r Sta tus w h e n y o u w a nt to de le te a ll m e s s a g e s in th e Cy to m e te r S ta tu s Me s s a g e s s c r e e n.

Fig ure 6.14 Cy tom eter Sta tus Messa g es Wind ow , Ex a m p le

Cy tom eter.log File

In th e c y to m e te r.lo g file , a ll m e s s a g e s a p p e a r fr o m th e la s t 30 da y s u nle s s th e y w e r e m a nu a lly c le a r e d. R e fe r to E R R O R ME S S A G E CY T O ME T E R .LO G F ILE fo r de ta ils . S e e F ig u r e 6.15.

PN 624923B 6-1 3

TROUBLESHOOTINGER R OR M ESSA G E CY TOM ETER .L OG F IL E 6

Fig ure 6.15 Ex a m p le of Error Messa g es in Cy tom eter.log File

6.4 ERROR MESSAGE CY TOMETER.LOG FILET h e Cy to m e te r e r r o r m e s s a g e s a r e lo c a te d in th e c y to m e te r.lo g file .

N ote: E r r o r m e s s a g e s o v e r 30 da y s o ld a r e a u to m a tic a lly r e m o v e d fr o m th e c y to m e te r.lo g file a nd p la c e d into th e c y to m e te r a rc h iv e .lo g file .

How to Access th e Cy tom eter.log File

From CX P Softw a re

r P r e s s Ý+L. S e e F ig u r e 6.15.

O R ,

r Cy tom eter tt Cy tom eter Log tt View Log . S e e F ig u r e 6.15.

From Wind ow s Desk top

1. Sta rt tt Prog ra m s tt Accessories tt Wind ow s Ex p lorer tt My Com p uter tt (C:) d riv e tt CX P.

2. Hig h lig h t cy tom eter.log .

3. D o u b le c lic k o n cy tom eter.log to o p e n. S e e F ig u r e 6.15.

PN 624923B6-1 4

TROUBLESHOOTINGER R OR M ESSA G E CY TOM ETER .L OG F IL E

Cy tom eter.Log Entry Descrip tion

A ll c y to m e te r.lo g e r r o r m e s s a g e e ntr ie s a r e p o s te d in c h r o no lo g ic a l o rde r. E a c h e r r o r m e s s a g e e ntr y c o ns is ts o f fo u r line s ( s e e F ig u r e 6.15) :

r D a te a nd tim e w h e n th e e r r o r m e s s a g e o c c u r r e d

r 18 -dig it nu m b e r - fo r S e r v ic e u s e o nly

r F o u r-dig it nu m b e r - fo r S e r v ic e u s e o nly

r E r r o r m e s s a g e te x t. S e e T a b le 6.2 fo r a lis t o f m e s s a g e s a nd o p e r a to r a c tio ns .

How to Sea rch th e Cy tom eter.log File

T o s e a rc h fo r a s p e c ific w o rd o r p h r a s e lis te d a ny w h e r e in th e c y to m e te r.lo g file :

1. Ed it tt Find ...

2. T y p e in th e w o rd o r p h r a s e y o u w a nt to find ( E x a m p le : Waste) .

3. T h e dir e c tio n o f th e s e a rc h : U p o r D o w n.

4. Find Nex t a nd th e ne x t o c c u r r e nc e o f th e w o rd in th e e r r o r lo g is h ig h lig h te d.

5. R e p e a t s te p 4 a s ne e de d o r u ntil C annot find “ X X X X X ” a p p e a r s .

Oth er Functions Av a ila b le

He re a r e s o m e o f th e m o r e o fte n u s e d fu nc tio ns a v a ila b le fr o m th e c y to m e te r.lo g file p u ll do w n m e nu s : File, Ed it, Form a t, a nd Help :

Print

T o p r int th e c y to m e te r.lo g file : File tt Print.

Ch a ng e Font

T o c h a ng e th e fo nt: Form a t tt Font, a nd s e le c t th e ne w fo nt.

Find Help Top ics

T o find He lp o n o th e r to p ic s : Help tt Help Top ics.

Cy tom etera rch iv e.log File

Th e c y to m e te r a rc h iv e .lo g file c o nta ins e r r o r m e s s a g e s th a t w e r e a u to m a tic a lly m o v e d fr o m th e c y to m e te r.lo g file a fte r 30 da y s . T h is lo g is fo r S e r v ic e u s e .

PN 624923B 6-1 5

TROUBLESHOOTINGER R OR M ESSA G ES TA B L E 6

6.5 ERROR MESSAGES TABLET a b le 6.2 lis ts th e e r r o r m e s s a g e s in a lp h a b e tic a l o rde r, w ith th e ir c a u s e a nd w h a t to do a b o u t th e m . T h e s e a r e th e e r r o r m e s s a g e s p r o du c e d b y th e Cy to m e te r.

Ca ll y o u r B e c k m a n Co u lte r R e p r e s e nta tiv e if:

r T h e r e c o m m e nde d a c tio n do e s no t s o lv e th e p r o b le m .

r Y o u ne e d h e lp .

Ta b le 6.2 Error Messa g es

Messa g e Ca use Action

Cle a n s e L e v e l Er ro r L ow cleaning ag ent. Fill cleaning ag ent container. R efer to FIL L T H E CL E ANING AG E NT CO NT AINE R .

Cleans e s ens or failed if th e cleaning ag ent container is full b ut th is error mes s ag e d is p lay ed .

Call y our Beckman Coulter R ep res entative.

Cle a n s e L e v e l W a r n in g L ow cleaning ag ent. Fill cleaning ag ent container. R efer to FIL L T H E CL E ANING AG E NT CO NT AINE R .

Cleans e s ens or failed if th e cleaning ag ent container is full b ut th is error mes s ag e d is p lay ed .


D a ta R a te W a r n in g T h e d is criminated d ata rate exceed ed 6,000 cells p er s econd .

1 . D ilute th e s amp le or ch ang e th e d is criminator s etting .

2. Ch eck th at th e s h eath fluid container cap is tig h tened .

3. If p rob lem continues , R E PL ACE T H E S H E AT H FL U ID FIL T E R .

D a ta Str e a m W a r n in g D ata acq uis ition h ard w are error 1 . R eb oot th e comp uter and res tart th e Cy tometer.

2. If p rob lem continues , call y our Beckman Coulter R ep res entative.

H o s t Co m m a n d In v a lid T h e Cy tometer s oftw are could not p roces s th e req ues ted command from th e W orks tation.

1 . R eb oot th e comp uter and res tart th e Cy tometer.


L a s e r Cu r r e n t Er ro r T h e amount of current req uired b y th e las er is outs id e th e exp ected rang e.


L a s e r P o w e r Er ro r T h e amount of las er lig h t p ow er is outs id e th e ins trument’s op erating rang e.


L a s e r R e g u la tio n W a r n in g T h e las er lig h t p ow er is fluctuating . Call y our Beckman Coulter R ep res entative.

L a s e r Sta rt Er r o r T h e las er failed to s tart w ith in 1 20 s econd s .

1 . Pow er th e ins trument off th en on.


PN 624923B6-1 6

TROUBLESHOOTINGER R OR M ESSA G ES TA B L E

M CL Ca r o u s e l In /Ou t Er r o r M CL carous el d id not move w h en req ues ted to d o s o.

1 . Ch eck th at th ere is no ob vious ob s truction (s amp le tub e) in th e M CL area.

2. Ch eck th at th e 30 p s i g ag e on th e p ow er s up p ly is okay .

3. If no ob s truction is found , call y our Beckman Coulter R ep res entative.

M CL Ca r o u s e l L a b e l Er r o r M CL carous el d id not read carous el b ar-cod e lab el.

1 . Ch eck th at th e b ar-cod e lab el is not torn or w ritten on.

2. T ry us ing anoth er carous el.


M CL Ca r o u s e l R o ta te Err o r M CL carous el d id not rotate correctly

Ch eck th at th ere is no ob vious ob s truction (s amp le tub e) in th e M CL area.

If no ob s truction is found , call y our Beckman Coulter R ep res entative.

M CL CP U Err o r M CL h ard w are error. Call y our Beckman Coulter R ep res entative.

M CL D o o r Op e n Er r o r M CL cover is op en w h ile th e M CL is in us e.

Clos e th e M CL cover.

M CL D o o r Op e n W a r n in g M CL cover is op en w h ile th e M CL is in us e.

Clos e th e M CL cover.

M CL EP R OM Err o r M CL h ard w are error. Call y our Beckman Coulter R ep res entative.

M CL Er r o r M CL h ard w are error. Call y our Beckman Coulter R ep res entative.

M CL In te rlo c k Er ro r T h e M CL Interlock is op en w h ile th e M CL is in us e.


M CL P a r a lle l R e c v Er r o r An M CL communications error. Call y our Beckman Coulter R ep res entative.

M CL P a r a lle l Se n d Er ro r An M CL communications error. Call y our Beckman Coulter R ep res entative.

M CL P r o b e U p /D o w n Err o r S h ort circuit. Ch eck if a circuit b reaker ind icator is d ark. S ee R E S E T T H E CIR CU IT BR E AK E R S .

M CL s amp le p rob e d id not move w h en req ues ted to d o s o.


M CL P r o g r a m L o a d Err o r An M CL h ard w are error. Call y our Beckman Coulter R ep res entative.

M CL R A M Er r o r An M CL h ard w are error. Call y our Beckman Coulter R ep res entative.

M CL R e c fr o m F a u lt Er r o r An M CL communications error. Call y our Beckman Coulter R ep res entative.

M CL R e c e iv e Tim e o u t Er r o r An M CL communication error. Call y our Beckman Coulter R ep res entative.

M CL R e q u e s t Er r o r Communication error b etw een th e Cy tometer and th e M CL .

1 . Pow er th e ins trument off th en on.


M CL Se r ia l 1 R e c v Er r o r An M CL communications error. Call y our Beckman Coulter R ep res entative.

M CL Se r ia l 1 Se n d Er ro r An M CL communications error. Call y our Beckman Coulter R ep res entative.

Ta b le 6.2 Error Messa g es (Continued)


PN 624923B 6-1 7


M CL Se r ia l 2 R e c v Er r o r An M CL communications error. Call y our Beckman Coulter R ep res entative.

M CL Se r ia l 2 Se n d Er ro r An M CL communications error. Call y our Beckman Coulter R ep res entative.

M CL Tr a n s m it Tim e o u t Er r o r An M CL communication error. Call y our Beckman Coulter R ep res entative.

M CL Tu b e D is p la c e d Er ro r A tub e is p os itioned incorrectly in th e M CL .


M CL Tu b e J a m Er r o r A tub e is s tuck in th e M CL . Call y our Beckman Coulter R ep res entative.

M CL Tu b e L o a d Err o r T h ere is a b ad s eal b etw een th e M CL s amp le p rob e g uid e and s amp le tub e.

Put s amp le into anoth er tub e.

M CL Tu b e P o s itio n Er r o r T h e M CL d id not rotate to th e correct tub e p os ition or th ere w as an error read ing th e p os ition b ar cod e on th e carous el.


M CL Tu b e U p /D o w n Er r o r U nab le to load or unload th e s amp le tub e from th e M CL s amp ling p os ition.

1 . Ch eck th at th e lab els on th e s amp le tub es are s ecure and are not ad h ering to th e w alls of th e carous el.

2. Ch eck th at th ere is no crack in th e s amp le tub e.


Sa m p le P r e s s u r e Er ro r T h ere may b e a leak caus ed b y a b ad s amp le tub e or a b ad s amp le h ead .

Ins p ect s amp le tub e and s amp le h ead for d amag e. Ch ang e as req uired . S ee R E PL ACE T H E M CL S AM PL E H E AD .

Bad reg ulator or tub ing . Call y our Beckman Coulter R ep res entative.

Se n s o rs Stre a m W a r n in g T h ere w as an error in th e communication of th e ins trument op erating d ata p acket from th e acq uis ition h ard w are to th e Cy tometer s oftw are.

1 . R eb oot th e comp uter and res tart th e Cy tometer.


Sh e a th D r a w e r Er ro r T h e s h eath fluid (reag ent) d raw er is op en.

Clos e th e reag ent d raw er comp letely .

Sh e a th L e v e l Er r o r T h ere is not enoug h s h eath fluid for furth er s amp le analy s is .

Fill s h eath fluid container. S ee FIL L T H E S H E AT H FL U ID CO NT AINE R .

Sh e a th L e v e l W a r n in g T h e firs t time th is mes s ag e is d is p lay ed , th ere is enoug h s h eath fluid for ab out 5 minutes of s amp le analy s is .

Fill th e s h eath fluid container. S ee FIL L T H E S H E AT H FL U ID CO NT AINE R .

If th e s h eath fluid container is full, th e s h eath fluid s ens or failed .




PN 624923B6-1 8

TROUBLESHOOTINGER R OR M ESSA G ES TA B L E

Sh e a th P r e s s u r e Er r o r T h e s h eath fluid p res s ure is outs id e th e s y s tem’s op erating rang e.

1 . Ch eck s h eath fluid and cleaning ag ent container cap s for tig h tnes s .

2. If p rob lems continues , call y our Beckman Coulter R ep res entative.

3. If p rob lem continues , R E PL ACE T H E S H E AT H FL U ID FIL T E R .

Sta te M a c h in e F a ilu r e Cy tometer s oftw are failure. 1 . Pow er th e ins trument off th en on.

2. If p rob lems continues , call y our Beckman Coulter R ep res entative.

Sy s te m P r e s s u r e Er ro r Pres s ure line is not connected b etw een th e p ow er mod ule and th e Cy tometer.

Connect th e p res s ure line.

T h e p res s uriz ed air s up p ly is outs id e th e s y s tem’s op erating rang e.

1 . G o to th e R ead y S tate, th en ch eck th at th e s y s tem p res s ure is 30 p s i.

2. R un a s amp le and monitor th e s y s tem p res s ure. If th e s y s tem p res s ure d rop s b elow th e rang e s p ecified in th e ins trument manual, th en call y our Beckman Coulter R ep res entative.

S h ort circuit. Ch eck if a circuit b reaker ind icator is d ark. S ee R E S E T T H E CIR CU IT BR E AK E R S .

Sy s te m Va c u u m Er r o r L iq uid in th e vacuum trap . Ch eck th at th e vacuum trap (on th e front of th e Pow er S up p ly ) is tig h t and is les s th an 1 /4 full of fluid . If it is more full, emp ty it (s ee CL E AN T H E V ACU U M T R AP).

V acuum line is not connected b etw een th e p ow er mod ule and th e Cy tometer.

Connect th e vacuum line at th e b ack of th e ins trument.

H ard w are p rob lem. Call y our Beckman Coulter R ep res entative.

Te m p e r a tu r e Err o r T emp erature ins id e th e Cy tometer is ab ove 60° C.

1 . Clean y our air filters . S ee CL E AN T H E AIR FIL T E R S .


Te m p e r a tu r e Se n s o r F a ilu r e T emp erature s ens or failure. Call y our Beckman Coulter R ep res entative.

Te m p e r a tu r e W a r n in g T emp erature ins id e Cy tometer is ab ove 50° C.

1 . Clean y our air filters . S ee CL E AN T H E AIR FIL T E R S .


Tra n s p u te r 1 Er r o r Cy tometer s oftw are error. At th e W orks tation turn th e s y s tem off, and th en turn it b ack on.

Tra n s p u te r 2 Er r o r Cy tometer s oftw are error. At th e W orks tation turn th e s y s tem off, and th en turn it b ack on.



PN 624923B 6-1 9


Tra n s p u te r L in k Er r o r A Cy tometer comp uter communications error.


Tra n s p u te r L in k W a r n in g A Cy tometer comp uter communications error.


U n k n o w n Str e a m W a r n in g An unid entified d ata p acket communication error occurred b etw een th e acq uis ition h ard w are and th e Cy tometer s oftw are.

1 . R ecord an entry in th e Action L og S h eet.

2. R eb oot th e comp uter and res tart th e Cy tometer.


Va c u u m Ch a m b e r W a r n in g T h e liq uid level in th e vacuum ch amb er is too h ig h .

1 . Ch eck th at th e vacuum trap (on th e front of th e Pow er S up p ly ) is tig h t and is les s th an 1 /4 full of fluid . If it is more full, emp ty it (s ee CL E AN T H E V ACU U M T R AP).


W a s te B a c k p r e s s u r e Er r o r T h e filter on th e w as te container vent line is p rob ab ly w et, clog g ed , or d is connected .

1 . Ch eck th e w as te vent filter for th e p res ence of liq uid and a p rop er connection.


W a s te L e v e l Er r o r T h ere is not enoug h emp ty volume in th e w as te container for furth er s amp le analy s is .

E mp ty th e w as te container. S ee E M PT Y T H E W AS T E CO NT AINE R .

W a s te L e v e l W a r n in g T h ere is enoug h emp ty volume in th e w as te container for ab out 5 minutes of s amp le analy s is .

E mp ty th e w as te container. S ee E M PT Y T H E W AS T E CO NT AINE R .

If th e w as te container is not full, th e w as te s ens or failed .




PN 624923B6-20

TROUBLESHOOTINGL EVEL SENSE IND ICA TOR S

6.6 LEVEL SENSE INDICATORS

Sh ea th Low

W h e n th e Sh ea th Low indic a to r ( s e e F ig u r e 6.16) a p p e a r s :

r D u r ing s a m p le a na ly s is , y o u h a v e 5 m inu te s to finis h a na ly z ing th e c u r r e nt s a m p le .

r Y o u c a nno t a na ly z e s a m p le s o r u s e th e ins tr u m e nt u ntil th e s h e a th flu id c o nta ine r is fille d.

r F ILL T HE S HE A T H F LU ID CO N T A IN E R .

Fig ure 6.16 Sh ea th Low Ind ica tors

Wa ste Full

W h e n th e Wa ste Full indic a to r ( s e e F ig u r e 6.17 ) a p p e a r s :

r D u r ing s a m p le a na ly s is , y o u h a v e 5 m inu te s to finis h a na ly z ing th e c u r r e nt s a m p le .

r Y o u c a nno t a na ly z e s a m p le s o r u s e th e ins tr u m e nt u ntil th e w a s te c o nta ine r is e m p tie d.

r E MP T Y T HE W A S T E CO N T A I N E R .

Fig ure 6.17 Wa ste Full Ind ica tors

6.7 CY TOMETER CONTROL WINDOW CANNOT BE VIEWEDIf th e Cy to m e te r Co ntr o l w indo w c a nno t b e v ie w e d ( it is h idde n b e h ind th e W indo w s T a s k b a r a t th e b o tto m o f th e s c r e e n) , th e A u to h ide fe a tu r e is c u r r e ntly s e le c te d. Ch a ng e th e W indo w s s e tting s to de s e le c t th e A u to h ide fe a tu r e a nd k e e p th e Cy to m e te r Co ntr o l w indo w in v ie w .

T o de s e le c t th e A u to h ide fe a tu r e :

b

b

PN 624923B 6-21

TROUBLESHOOTINGCY TOM ETER CONTR OL W IND OW CA NNOT B E VIEW ED 6

1 Sta rt tt Setting s tt Ta sk b a r & Sta rt

Menu....

2 O n th e Genera l ta b , Auto h id e

a nd de s e le c t Alw a y s on top .

.

3 T h e Cy to m e te r Co ntr o l w indo w s h o u ld

no w b e p a r tia lly in v ie w .

4 D r a g th e Cy to m e te r Co ntr o l w indo w

into fu ll v ie w .

PN 624923B6-22

TROUBLESHOOTINGCY TOM ETER CONTR OL W IND OW CA NNOT B E VIEW ED

PN 624923B A-1

AAHOW TO...

A.1 SOFTWARE PROCEDURESF o r O nline h e lp , c lic k o n th e link s b e lo w to g o to th e de ta ile d p r o c e du r e .

Th ese p roced ures a re in th e Sy stem Ov erv iew ch a p ter in th e Instructions For Use Ma nua l.

r CR E A T IN G P R O T O CO LS

r CR E A T IN G R E G I O N S

r CR E A T IN G G A T E S

r CR E A T IN G F LO W P A G E S

r CR E A T IN G P A N E LS

r CR E A T IN G W O R K LIS T S

Th ese p roced ures a re in th e Qua lity Control ch a p ter in th e Instructions For Use Ma nua l.

r A u to S e tu p I I W iz a rd

r A p p lic a tio n D e finitio n W iz a rd ( de fine a ll y o u r a p p lic a tio ns )

r A u to S e tu p S c h e du le r ( m u ltip le a p p lic a tio n A u to S e tu p )

Th ese p roced ures a re in th e Using CX P Softw a re ch a p ter in th e Reference Ma nua l.

r B a tc h A u to MA T O R

r O v e r la y His to g r a m P lo ts

r A dv a nc e d Co lo r P r e c e de nc e

r Lis tm o de Co m p e ns a tio n

r S e tting a CA L R e g io n

A.2 HARDWARE PROCEDURESF o r O nline h e lp , c lic k o n th e link s b e lo w to :

r V ie w a s h o r t v ide o o f th e p r o c e du r e o r

r G o to th e de ta ile d p r o c e du r e .

Th ese p roced ures a re in th e Sp ecia l Proced ures Ma nua l.

r O p e n/r e m o v e th e ins tr u m e nt c o v e r s - v ide o o r p r o c e du r e .

r Cle a n th e MCL s a m p le h e a d a nd th e s a m p le p r o b e - v ide o o r p r o c e du r e .

r R e p la c e th e s a m p le p r o b e a nd s a m p le p ic k u p tu b ing - v ide o o r p r o c e du r e .

r R e p la c e th e MCL s a m p le h e a d - v ide o o r p r o c e du r e

r R e p la c e th e o p tic a l filte r p la te - v ide o o r p r o c e du r e .

r R e p la c e a filte r in th e o p tic a l filte r p la te - v ide o o r p r o c e du r e

PN 624923B A-2

HOW TO...H A R D W A R E P R OCED U R ES

PN 624923B B-1

BBBAR-CODE SPECIFICATIONS

B.1 BAR-CODE SAMPLE IDENTIFICATIONB a r-c o de s y m b o ls a r e a h ig h ly a c c u r a te a nd e ffic ie nt p r o c e du r e fo r ide ntify ing a nd p r o c e s s ing la b o r a to r y s a m p le s . B e c k m a n Co u lte r ins tr u m e nts u s e fo u r b a r-c o de s y m b o lo g ie s ( ty p e s ) to ide ntify s p e c im e ns :

r Co de 128

r Co de 39®

r Co da b a r

r Inte r le a v e d 2-o f-5.

T h e b a r- c o de r e a de r s e ns e s th e diffe r e nc e b e tw e e n e na b le d b a r-c o de s y m b o lo g ie s in a r u n.

Fig ure B.1 Ba r-Cod e La b el

B.2 CORRECT PLACEMENT OF THE BAR-CODE LABELT h e b a r- c o de la b e l m u s t b e p la c e d a m inim u m o f 25.4 m m ( 1.0 in.) fr o m th e b o tto m o f th e tu b e . R e fe r to F ig u r e B .2.

IMPORTANT A mis read lab el can caus e one s amp le ID to b e read as anoth er s amp le ID . T h e lab oratory ’s p roces s for p rinting , p lacing , and meeting all b ar-cod e s p ecifications is imp ortant to ach ieve h ig h ly accurate read ing . Follow th e b ar-cod e s p ecifications to avoid inaccurate read ing of th e b ar-cod e lab el.

b Q u ie t z o ne

c B a r- c o de s y m b o l

d S a m p le I D

AB C1 2 3 4 5 6 7 8 9

b

c

d

0231001A

PN 624923BB-2

BAR-CODE SPECIFICATIONSB A R -COD E L A B EL SP ECIF ICA TIONS

Fig ure B.2 Ba r-Cod e La b el Pla cem ent

P u t la b e ls o n th e tu b e s s o th a t th e b a r s fo llo w o ne a no th e r in a v e r tic a l s e q u e nc e . R e fe r to F ig u r e B .2. T h e b a r-c o de r e a de r s c a ns th e tu b e v e r tic a lly . D o no t tilt th e la b e l m o r e th a n ± 7 .5 de g r e e s fr o m th e a x is o f th e tu b e .

P u t th e tu b e s in th e c a r o u s e l s o th a t th e b a r- c o de s y m b o ls a r e v is ib le th r o u g h th e s lo ts in th e fr o nt o f th e c a r o u s e l. W h e n v ie w e d a t e y e le v e l, th e fu ll s y m b o l, inc lu ding th e q u ie t z o ne s , m u s t b e v is ib le th r o u g h th e s lo t a nd a b o v e th e b o tto m o f th e c a r o u s e l.

N ote: T h e F C 50 0 r o ta te s th e tu b e a s ne e de d s o th e b a r-c o de la b e l c a n b e r e a d.

B.3 BAR-CODE LABEL SPECIFICATIONS

T h e q u a lity o f th e b a r- c o de s y m b o l a nd th e la b e l is im p o r ta nt fo r a c c u r a te r e a ding . F o r h ig h a c c u r a c y , u s e la b e ls th a t m e e t a ll o f th e s p e c ific a tio ns .

W h e n p o s s ib le , p r int th e s a m p le I D o n th e la b e l in a lp h a nu m e r ic c h a r a c te r s s o th e o p e r a to r c a n m a nu a lly e nte r th e b a r- c o de info r m a tio n if th e b a r-c o de s y m b o l c a nno t b e r e a d.

La b el Siz e a nd Th ick ness

T h e le ng th o f th e la b e l m u s t b e le s s th a n 44.45 m m ( 1.7 5 in.) . T h e la b e l inc lu de s th e b a r- c o de s y m b o l a nd a m inim u m q u ie t z o ne o f 3.5 m m ( 0 .14 in.) a t e a c h e nd o f th e s y m b o l. R e fe r to F ig u r e B .3.

b 25.4 m m ( 1.0 in.) m inim u m

c 7 .5 de g r e e s

b

c

c

IMPORTANT A mis read lab el can caus e one s amp le ID to b e read as anoth er s amp le ID . T h e lab oratory ’s p roces s for p rinting , p lacing , and meeting all b ar-cod e s p ecifications is imp ortant to ach ieve h ig h ly accurate read ing . Follow th e b ar-cod e lab el s p ecifications to keep th e rate of mis read lab els to a minimum.

PN 624923B B-3

BAR-CODE SPECIFICATIONSB A R -COD E L A B EL SP ECIF ICA TIONS B

Fig ure B.3 Ba r-Cod e La b el Sp ecifica tions

T h e w idth o f th e b a r- c o de la b e l m u s t b e 5 m m ( 0 .2 in.) le s s th a n th e c irc u m fe r e nc e o f th e s a m p le tu b e .

La b e l th ic k ne s s , inc lu ding a dh e s iv e , m u s t b e 0 .0 9 m m ( 0 .0 0 36 in.) m a x im u m . T o ta l th ic k ne s s fo r a ll la b e ls a nd a dh e s iv e s p u t to g e th e r m u s t b e 0 .36 m m ( 0 .0 144 in.) m a x im u m .

Sy m b ol Dim ensions

T h e h e ig h t o f th e b a r- c o de s y m b o l m u s t b e 19.0 5 m m ( 0 .7 5 in.) m inim u m .

S e e T a b le B .2.

La b el a nd Print Qua lity

A ll b a r- c o de s y m b o ls m u s t a g r e e w ith th e A m e r ic a n Ide ntific a tio n Ma nu fa c tu r e r ’s ( A IM)

U nifo r m S y m b o lo g y S p e c ific a tio n.1

A ll b a r- c o de s y m b o ls m u s t b e p r inte d a t p r int q u a lity c la s s “ B ” o r b e tte r a s de fine d b y th e

A m e r ic a n N a tio na l S ta nda rds Ins titu te ( A N S I) .2 S e v e r a l fa c to r s a ffe c t p r int q u a lity :

r La b e ls m u s t b e c le a n, no t y e llo w e d, a nd u s e d b e fo r e th e e x p ir a tio n da te .

r P r int th e b a r- c o de s y m b o l o n m a te r ia l th a t is r e fle c tiv e a nd h a s a m a tte finis h . U s e a b a c k g r o u nd diffu s e r e fle c ta nc e o f 8 0 % o r m o r e fo r m a x im u m c o ntr a s t.

r T h e la b e ls m u s t no t h a v e de fe c ts s u c h a s s p o ts , line s , m is s ing s e c tio ns , c u ts , fo lds , o r de ns ity p r o b le m s .

r T h e b a r s in th e b a r- c o de s y m b o l m u s t b e w e ll-de fine d. E dg e s m u s t b e c o ns ta nt ( no t ir r e g u la r ) , s o th e b a r s a nd s p a c e s h a v e th e c o r r e c t w idth s fo r th e b a r- c o de s y m b o lo g y u s e d.

b Q u ie t z o ne s 3.5 m m ( 0 .14 in.) m inim u m

c B a r- c o de s y m b o l h e ig h t 19.0 5 m m ( 0 .7 5 in.) m inim u m

d B a r- c o de la b e l le ng th 44.45 m m ( 1.7 5 in.) m a x im u m

AB C1 2 3 4 5 6 7 8 9

b b

c

d0231002A

PN 624923BB-4

BAR-CODE SPECIFICATIONSB A R -COD E ER R OR R A TE

B.4 BAR-CODE ERROR RATE

T h e q u a lity o f th e b a r-c o de s y m b o l a nd th e la b e l is im p o r ta nt fo r a c c u r a te r e a ding . T o g e t th e h ig h e s t p o s s ib le a c c u r a c y o nly u s e la b e ls th a t m e e t a ll th e s p e c ific a tio ns de s c r ib e d fo r la b e ls a nd s y m b o ls . D e v ia tio ns fr o m th e s e s p e c ific a tio ns m a k e th e b a r c o de m o r e diffic u lt to r e a d a nd a llo w fo r a p o s s ib le inc r e a s e in th e e r r o r r a te .

T h e s y m b o lo g y a nd th e c o nfig u r a b le p a r a m e te r s th a t th e la b o r a to r y s e le c ts h a v e a n e ffe c t o n th e e r r o r r a te . Ce r ta in fe a tu r e s o f th e s y m b o lo g ie s a nd th e s e le c tio ns m a de b y th e la b o r a to r y h a v e a n im p o r ta nt e ffe c t o n th e a c c u r a c y o f th e b a r- c o de r e a ding s y s te m . In g e ne r a l:

r Co de 128 a nd Co de 39 a r e m o r e a c c u r a te a nd h a v e lo w e r e r r o r r a te s th a n Co da b a r o r Inte r le a v e d 2-o f-5.

r N CCLS r e c o m m e nds Co de 128 b e c a u s e o f its a c c u r a c y , c o m p a c t fo r m , a nd s e lf-c h e c k ing

c a p a b ilitie s .3

r A c h e c k s u m g r e a tly inc r e a s e s a c c u r a c y . U s e a c h e c k s u m w ith Inte r le a v e d 2-o f-5 a nd Co da b a r b e c a u s e th e y a r e le s s a c c u r a te s y m b o lo g ie s .

r S e le c t th e fix e d le ng th o p tio n, if a v a ila b le , b e c a u s e it is m o r e a c c u r a te th a n th e v a r ia b le le ng th o p tio n.

r T o k e e p la b e l a nd p r inting fla w s to a m inim u m , u s e a na r r o w e le m e nt o f m o r e th a n 0 .25 m m ( 0 .0 10 in.) .

Beck man Cou lter recommen d s the u se of:

r Cod e 128

r Check su m for all other symb olog ies

r Fix ed len g th cod e symb ols

r N arrow b ar siz es of 0.25 mm (0.010 in .) min imu m.

IMPORTANT A mis read lab el can caus e one s amp le ID to b e read as anoth er s amp le ID . W h enever p os s ib le us e a b ar-cod e s y mb olog y and config uration ch oices th at p rovid e th e mos t accurate b ar-cod e read ing .

PN 624923B B-5

BAR-CODE SPECIFICATIONSB A R -COD E SY M B OL OG IES B

B.5 BAR-CODE SY MBOLOGIESB e c k m a n Co u lte r ins tr u m e nts u s e fo u r b a r-c o de s y m b o lo g ie s fo r s p e c im e n ide ntific a tio n, s e e T a b le B .1.W ith in th e g iv e n s p e c ific a tio ns , th e MCL r e a de r a nd th e o p tio na l h a ndh e ld b a r- c o de r e a de r a u to m a tic a lly dis ting u is h th e fo llo w ing b a r c o de s :

Ta b le B.1 Ba r-Cod e Sy m b olog ies

Ba r-Cod e Ty p e Descrip tion

Cod e 1 28(als o know n as U S D -6)

r V ariab le leng th

r Alp h anumerics ; 1 07 ch aracter s et

r S elf-ch ecking

r Continuous cod e; interch aracter s p ace is p art of cod e s tructure for h ig h er d ens ity of cod e p er s q uare inch ; comp act b ar cod e

r Cod e 1 28 is recommend ed b y th e National Committee for Clinical L ab oratory S tand ard s (NCCL S ) for its accuracy , comp act form, and s elf-ch ecking cap ab ilities 3

r Cod e 1 28B - M aximum 8 alp h anumeric ch aracters

r Cod e 1 28C - M aximum 1 6 numeric ch aracters(T h e us e of 1 5 numeric ch aracters is invalid )

Cod e 39(als o know n as 3-of-9 and U S D -3)


r Includ es 43 d ata ch aracters ; 26 letters (up p ercas e A-Z ), 1 0 d ig its (0-9), s ix s y mb ols (. $ / + % -) and a s p ace

r S trong s elf-ch ecking p rop erties

r Ch ecks um

r D is crete cod e; w h ite s p aces are not p art of th is cod e

r M aximum 7 ch aracters(6 d ata ch aracters + 1 ch ecks um ch aracter).

Interleaved 2-of-5(als o know n as I2 of 5, U S D -1 , and U S D -1 .25)

r Numerics only

r Ch ecks um

r L ow er d ens ity of cod e p er s q uare inch ; long er lab el

r R eq uires an even numb er of d ig its to b e encod ed , a lead ing “ 0” mus t b e ad d ed if th e numb er count is od d

r Fixed 1 4 ch aracters(1 3 d ata ch aracters + 1 ch ecks um ch aracter).

Cod ab ar(als o know n as U S D -4 and NW 7 )


r Includ es 1 6 d ata ch aracters ; 1 0 d ig its (0-9), and s ix s y mb ols (. $ / + % -)

r H as s p ecific s tart and s top ch aracters w h ich lead to imp rovement in read ab ility

r Ch ecks um

r L ow er d ens ity of cod e p er s q uare inch ; long er b ar cod e

r M aximum 1 0 ch aracters(9 d ata ch aracters + 1 ch ecks um ch aracter).

PN 624923BB-6

BAR-CODE SPECIFICATIONSB A R -COD E L A B L ES

B.6 BAR-CODE LABLESA b a r c o de c o ns is ts o f b la c k line s ( b a r s ) a nd w h ite line s ( s p a c e s ) , w h ic h a r e c a lle d e le m e nts .

T h e r e a r e na r r o w e le m e nts ( N E ) a nd w ide e le m e nts ( W E ) . T h e b a r- c o de s y m b o lo g y de te r m ine s th e ir a r r a ng e m e nt.

T h e ins tr u m e nt s u p p o r ts p r e p r inte d la b e ls .

Ba r-Cod e La b el Op tica l Ch a ra cteristics a t 670 nm ± 10 %

r P r int Co ntr a s t S ig na l ( P CS ) : 8 0 % m inim u m .

r R e fle c tiv ity o f Me dia ( R W ) : 8 0 % m inim u m .

r R e fle c tiv ity o f Ink ( R B ) : 16% m a x im u m .

r N o s p o ts o r v o ids ; no ink s m e a r ing .

r E dg e r o u g h ne s s is inc lu de d in th e b a r a nd s p a c e to le r a nc e s .

B.7 MCL BAR-CODE READERT h e MCL u s e s a v is ib le -la s e r ty p e r e a de r c o nta ining a Cla s s II la s e r, o p e r a ting a t 67 0 nm , w ith a m a x im u m p o w e r o u tp u t o f 1 m W .

IMPORTANT S amp le mis id entification can occur from th e us e of incorrect, p oor q uality , d amag ed , d irty or imp rop erly p laced b ar-cod e lab els . Follow th e s p ecifications in th is s ection to create y our b ar-cod e lab els to p revent incorrect s amp le id entification. S ee als o Putting a Bar-Cod e L ab el on a S amp le T ub e.

Ta b le B.2 Cod e-Rela ted Sp ecifica tions

Cod eInterlea v ed 2-of-5* Cod a b a r* Cod e 39* Cod e 128*

Narrow element (NE ) w id th 0.01 0 in. ± 0.001 0.01 0 in. ± 0.001 0.01 0 in. ± 0.001 0.01 0 in. ± 0.001

W id e element/narrow element ratio (W E /NE )

3:1 N/A 3:1 N/A

Interch aracter g ap No 0.01 0 in. minimum _ NE No

D ata d ig its 1 4* * 1 to 1 0* * 1 to 7 * * 2 to 1 6

* S ee AIM U niform S y mb olog y S p ecification, R ev. 1 995 for d etailed s p ecification.* * Includ es ch ecks um ch aracter

P C SRW RB+

RW------------------------ 100%×=

PN 624923B B-7

BAR-CODE SPECIFICATIONSB A R -COD E D ECOD ER B

B.8 BAR-CODE DECODERT h e MCL s e nds a “ G S ” A S CII c h a r a c te r ( h e x a de c im a l 1D ) to th e de c o de r to s ta r t o p e r a tio n.

T h e de c o de r :

r T u rns th e r e a de r o n.

r D e c o de s info r m a tio n th a t c o m e s fr o m th e r e a de r.

r K e e p s th e r e a de r o n fo r u p to 4 s e c o nds .

r T u rns th e r e a de r o ff.

r S e nds th e de c o de d info r m a tio n ( o r no -r e a d m e s s a g e ) to th e MCL.

B.9 CHECKSUM ALGORITHMB e c k m a n Co u lte r s tr o ng ly r e c o m m e nds th e u s e o f b a r c o de c h e c k s u m s to p r o v ide a u to m a tic c h e c k s fo r r e a d a c c u r a c y .

IMPORTANT T o p revent incorrect id entification of s amp le tub es , d o not us e FNC1 , FNC4, and FS (h exad ecimal 1 C) ch aracters in y our b ar-cod e information.

IMPORTANT U s e of b ar cod es is an extremely accurate and effective meth od of p os itive p atient id entification. Certain features , s uch as ch ecks um d ig its , maximiz e accuracy in read ing Cod ab ar, Cod e 39 and Interleaved 2-of-5 lab els . In one s tud y , th e us e of ch ecks um d ig its d etected 97 % of mis read errors .

U s e ch ecks ums to p rovid e p rotection ag ains t occas ional mis read errors caus ed b y p rob lems s uch as d amag ed or mis ap p lied lab els . If y ou mus t us e b ar cod es w ith out ch ecks ums , Beckman Coulter recommend s th at y ou verify each b ar-cod e read ing to as s ure correct p atient id entification.

PN 624923BB-8

BAR-CODE SPECIFICATIONSCH ECK SU M A L G OR ITH M

PN 624923B R E FE R E NCE S -1

REFERENCES

1. A m e r ic a n Ide ntific a tio n Ma nu fa c tu r e r ’s g r o u p ( A IM) , U niform S y mbology S pecifications C ode 3 9 , Interleaved 2 of 5 , C odabar, a nd International S y mbology S pecifications C ode 1 2 8 . A N S I/A IM B C1, B C2, B C3, B C4, 1995. h ttp ://w w w .a im u s a .o r g

2. A m e r ic a n N a tio na l S ta nda rds Ins titu te ( A N S I) B ar C ode P rint Q uality G uidelines. X 3. 18 2-1990 ( R 20 0 0 ) . h ttp ://w w w .a ns i.o r g

3. N a tio na l Co m m itte e fo r Clinic a l La b o r a to r y S ta nda rds ( N CCLS ) , L aboratory A utomation: B ar C odes for S pecimen C ontainer Identification; A pproved S tandard. A U T O 2-A ( I S B N 1-56238 -360 -4) . h ttp ://w w w .nc c ls .o r g

PN 624923BR E FE R E NCE S -2

REFERENCES

PN 624923B IND E X -1

INDEX

Num erics3D a c c e le r a tio n

s e tting ne e de d o n v ide o c a rd, 1-123dfx v o o do o b a s e d v ide o c a rds

s e tting ne e de d fo r 3D a c c e le r a tio n, 1-123-o f-9

b a r -c o de , B -548 8 D L filte r

u s e d to s e p a r a te S S , 5-6S ee also filte r s

50 0 LP filte ru s e d to b lo c k la s e r lig h t, 5-7S ee also filte r s

Aa b o u t h e lp

lis ts CX P s o ftw a r e b u ild nu m b e r, 1-38a b s o lu te c o u nts

u s e F lo w -Co u nt flu o r o s p h e r e s , 3-1a c c e s s do o r , tu b e

lo c a tio n, 4-10A cquiring

a p p e a r s du r ing s a m p le c y c le , 4-4, 4-8 , 4-11a c q u is itio n

m o nito r w ith e v e nts /s e c c o u nte r, 4-4, 4-8 , 4-11r u nning s a m p le s in th e a u to m a tic m o de , 4-2r u nning s a m p le s in th e m a nu a l m o de , 4-5r u nning s a m p le s in th e s ing le tu b e m o de , 4-9

a c q u is itio n m a na g e rdr a g a nd dr o p fu nc tio ns , 1-19e nte r th e c a r o u s e l ID nu m b e r, 4-4, 4-7s k ip tu b e s in c a r o u s e l, 1-14to o lb a r

w o r k lis t p a ne l, 1-35w o r k lis t te s t, 1-36

A c tio n lo g s h e e tr e c o rd c le a ning , 2-11r e c o rd da ily s h u tdo w n a c tio ns , 2-11r e c o rd s ta r tu p c h e c k s , 2-8

a dju s tc o lo r c o m p e ns a tio n, 3-1g a in, 3-1h ig h v o lta g e , 3-1s y s te m p r e s s u r e , 2-6S ee also a dju s tm e nt p r o c e du r e s

a dju s tm e nt p r o c e du r e ss c h e du le , 6-11s y s te m p r e s s u r e , 2-6

a ir filte rda ily c h e c k , 2-6w h e n to c a ll s e r v ic e , 2-6

a lig nm e ntv e r ify , 3-1

a m p lific a tio nde s c r ip tio n, 5-10

A P CS ee also F lo w -Ch e c k 67 5 a nd F lo w -S e t 67 5

flu o r o s p h e r e sa p p lic a tio n

c o ntr o l, v e r ify , 3-1IV D , 1-1lis t, fo r th e ins tr u m e nt, 1-2

a p p lic a tio n de finitio nw iz a rd, 3-10

A r g o n la s e rde s c r ip tio n o f h o w its lig h t is h a ndle d, 5-7illu s tr a tio n, 5-5la s e r la b e ls , 6-5O N indic a to r, 1-4S ee also la s e r

a u to a dju s t dis a b le du s e w h e n r u nning a ne g a tiv e c o ntr o l, 3-7

a u to m o deS ee a u to m a tic m o de

a u to s h u tdo w nh o w to s e t u p , 2-14

a u to s ta r tu ph o w to s e t u p , 2-14

A u to G a te r e g io nc o nv e r t fr o m a p o ly g o na l r e g io n, 1-30c r e a ting , 1-28s to p c o u nt o n p lo t, 1-28

a u to m a te d s o ftw a r efe a tu r e s , 5-12

a u to m a ticm o de , r u nning , 4-2

a u to m a tic g a te c r e a tio n, 1-30a u to m a tic m o de

r u nning s a m p le s , 4-2A u to MA T O R

s e tu pdr a g a nd dr o p fu nc tio ns , 1-19

A u to S e tu pc o m p e ns a tio n tu b e s , 3-3p a ne ls

c o ntr o ls u s e d, 3-1p r e p a r e fo r, 3-2

PN 624923BIND E X -2

INDEX

r u nning a ne g a tiv e c o ntr o l, 3-7v e r ific a tio n tu b e s , 3-3w iz a rd, r u nning , 3-5

a u to s e tu ps c h e du le r, 3-4w iz a rd

A u to S e tu p IIs a v e s e tting s , 3-9

a u to s ta nda rdiz a tio nS ee A u to S e tu p II

A U X p a r a m e te rde s c r ip tio n, 5-12dis p la y indic a to r, 1-5no t a v a ila b le w ith F L5, 5-12w h e n to u s e , 5-12

A waiting S amplea p p e a r s du r ing s a m p le c y c le , 4-4, 4-8 , 4-11a p p e a r s du r ing s ta r tu p c y c le , 2-5r u n flu o r o s p h e r e s , 3-5

Bb a r -c o de

3-o f-9, B -5a c c e p ta b le , B -5Co da b a r, B -5Co de 128 , B -5Co de 39, B -5e r r o r r a te , B -4I2-o f-5, B -5Inte r le a v e d 2-o f-5, B -5N W 7 , B -5s p e c ific a tio ns , B -1s y m b o lo g ie s , B -5U S D -1, B -5U S D -1.25, B -5U S D -3, B -5U S D -4, B -5U S D -6, B -5u s e o f c h e c k s u m , B -7

b a r -c o de de c o de rde s c r ip tio n, B -7

b a r -c o de la b e lsb a s ic o p e r a ting te c h niq u e s , 1-13c o r r e c t p la c e m e nt, B -1la b e l q u a lity , B -3la b e l s iz e , B -2la b e ling s a m p le tu b e s , 1-13o p tic a l c h a r a c te r is tic s , B -6o r ie nta tio n in c a r o u s e l, 1-14

p r int q u a lity , B -3s p e c ific a tio ns , B -2, B -6s y m b o l dim e ns io ns , B -3th ic k ne s s , B -2ty p e s r e a d b y th e MCL, B -5u s e d o n s a m p le c a r o u s e l, 5-1

b a r -c o de r e a de ra c c e p ta b le b a r c o de s , B -5la s e r la b e ls , 6-9MCL, s p e c ific a tio ns , B -6

b a s ic o p e r a ting te c h niq u e s , 1-13b e a ds

S ee flu o r o s p h e r e sb e a m s h a p ing

la s e r, 5-3b le a c h

u s e d in s h u tdo w n c le a ning , 2-11b lo o d

r u n a s a c o ntr o l, 3-1b lu e la s e r

S ee A r g o n la s e rb u tto ns

T a s k b a r, 1-8S ee also ic o ns , S ta r t b u tto n

Cc a lc u la ting

c o m p e ns a tio n c o e ffic ie nts , 3-7c a r o u s e l, s a m p le

b a r -c o de la b e ls , 5-1de s c r ip tio n, 5-1h o m e p o s itio n, 1-15lo a ding s a m p le tu b e s , 1-14p o s itio n 10 u s e d fo r s ing le tu b e m o de , 4-10p u tting a c a r o u s e l in th e MCL, 1-14r e m o v ing a c a r o u s e l fr o m th e MCL, 1-16S ee also MCL

c a r r yr e g io ns fr o m p r e v io u s p r o to c o l, 1-34

CD R Hr e q u ir e d la b e ls , 6-1

c e ll illu m ina tio nde s c r ip tio n, 5-4

c h a r a c te r is tic s , p e r fo r m a nc e , 1-38c h e c k

a ir filte r, 2-6p r e s s u r e g a u g e , 2-6v a c u u m filte r, 2-6v a c u u m g a u g e , 2-6

PN 624923B IND E X -3

INDEX

v a c u u m tr a p , 2-8w a te r tr a p , 2-6

c h e c k s u m a lg o r ith mCo da b a r, B - 7Co de 39, B - 7Inte r le a v e d 2-o f-5, B - 7

c le a nings c h e du le , 6-11

c le a ning a g e ntc h e c k le v e l a t s ta r tu p , 2-3c o nta ine r , w h e n to c le a n, 6-11

c le a ning p r o c e du r e ss c h e du le , 6-11s u r fa c e a r e a s du r ing s h u tdo w n, 2-11

C leanse L evel E rror, 6-15C leanse L evel Warning, 6-15Co da b a r

b a r -c o de , B -5Co da b a r b a r c o de

c h e c k s u m a lg o r ith m , B -7Co de 128

b a r -c o de , B -5Co de 39

b a r -c o de , B -5Co de 39 b a r c o de

c h e c k s u m a lg o r ith m , B -7c o e ffic ie nts

c a lc u la ting c o m p e ns a tio n, 3-7c o lo r c o m p e ns a tio n

a dju s t, 3-1c a lc u la ting c o e ffic ie nts , 3-7

c o lo r r e s o lu tio nh o w to s e t, 1-11m o nito r, 1-11

c o m p e ns a tio nc a lc u la ting c o e ffic ie nts , 3-7S ee also c o lo r c o m p e ns a tio n

c o m p e ns a tio n c o e ffic ie ntsc o m p e ns a tio n tu b e s , 3-7v e r ific a tio n tu b e s , 3-8

c o m p e ns a tio n tu b e sA u to S e tu p , 3-3p r e p a r e , 3-3p r o c e s s , 3-7u s e d to c a lc u la te c o m p e ns a tio n c o e ffic ie nts , 3-7

c o m p u te rfr o nt v ie w , 1-7

Co ntr o l P a ne lde s c r ip tio n, 1-10h o w to a c c e s s , 1-10

illu s tr a tio n, 1-10W indo w s , 1-10

c o ntr o lsa nd indic a to r s , Cy to m e te r, 1-3fo r A u to S e tu p p a ne ls , 3-1p r e p a r e fo r A u to S e tu p , 3-2Q C p r o c e s s e s u s e d fo r, 3-1r u nning , 3-1r u nning A u to S e tu p , 3-5r u nning F lo w -Ch e c k , 3-2w h o le b lo o d, 3-1S ee also indiv idu a l Q C m a te r ia l na m e s

c o nv e ntio nsu s e d in th e m a nu a l, x iv

c o r r e c t s e tting sv e r ify , 3-1

c r e a tingA u to G a te r e g io ns , 1-28F lo w P A G E , 1-31line a r r e g io ns , 1-26m u ltip le line a r r e g io ns , 1-27p o ly g o na l r e g io ns , 1-22q u a dr a nt r e g io ns , 1-24r e c ta ng u la r r e g io ns , 1-23

c r o s s - c y lindr ic a l le ns e sb e a m s h a p ing , 5-3

CVfo r m u la , 5-16

CX P s o ftw a r ea u to m a te d fe a tu r e s , 5-12b u ild nu m b e r, 1-38s h o r tc u ts , 1-17s h o r tc u ts o n S ta r t b u tto n m e nu , 1-8s ig ning o n, 2-5v e r s io n info r m a tio n, 1-38w h e r e e r r o r m e s s a g e s a p p e a r, 6-11S ee also s o ftw a r e

Cy to -Co m p Ce llsQ C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

Cy to m e te ra u to s h u tdo w n, 2-14a u to s ta r tu p , 2-14c o ntr o ls a nd indic a to r s , 1-3fr o nt v ie w , 1-3indic a to r p a ne ls , 1-4la s e r la b e ls , 6-2, 6-3, 6-4, 6-7r e a dy a nd la s e r o n indic a to r s , 1-4s a m p le flo w r a te indic a to r s , 1-5s e ns o r s , 5-8


INDEX

s ig na l a m p litu de indic a to r s , 1-5w h e n to s h u tdo w n, 2-9

Cy to m e te r A u to S ta r tu p , 2-14Cy to m e te r Co ntr o l

dr a g a nd dr o p fu nc tio ns , 1-19Cy to m e te r S ta tu s W indo w

de s c r ip tio n, 6-12illu s tr a tio n, 6-12S ee also e r r o r m e s s a g e s

c y to m e te r .lo g filede s c r ip tio n, 6-13illu s tr a tio n, 6-13S ee also e r r o r m e s s a g e s

c y to m e te r a r c h iv e .lo g filede s c r ip tio n, 6-14S ee also e r r o r m e s s a g e s

Cy to -T r o l Co ntr o l Ce llsQ C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

Dda ily p r o c e du r e s

b e fo r e y o u b e g in, 2-1P r inte r c h e c k s , 2-8Q C p r o c e du r e s , 3-2r e c o rd s h u tdo w n a nd c le a ning , 2-11r e c o rd s ta r tu p c h e c k s , 2-8s e t u p a u to s h u tdo w n, 2-14s e t u p a u to s ta r tu p , 2-14s h u tdo w n, 2-9s ta r tu p , 2-1s ta r tu p p r o c e du r e , 2-1

da ily s ta r tu pb e fo r e y o u b e g in, 2-1S ee also s ta r tu p

D ata R ate Warning, 6-15da ta s to r a g e

de s c r ip tio n, 5-15D ata S tream Warning, 6-15de le te

R e c y c le B in, 1-9D e s k to p

W indo w s , 1-8D e s k to p Me nu B a r

illu s tr a tio n, 1-8dia lo g b o x

de s c r ip tio n, x vdic h r o ic filte r s

S ee filte r s

dis p la y p a ne lflo w r a te indic a to r s , 1-5illu s tr a tio n, 1-4la s e r o n indic a to r, 1-4r e a dy indic a to r, 1-4s ig na l a m p litu de indic a to r s , 1-5

do c u m e nta tio n fo r y o u r ins tr u m e nta b o u t y o u r ins tr u c tio ns fo r u s e m a nu a l, x iiic o nv e ntio ns u s e d, x ivintr o du c tio n to y o u r m a nu a ls , x iiiu s ing th e m a nu a ls , x iii

D o c u m e ntsW indo w s , 1-9

do o r , tu b e a c c e s slo c a tio n, 4-10

do u b le t dis c r im ina tio nu s e th e A U X p a r a m e te r, 5-12

dr a g a nd dr o pa c q u is itio n m a na g e r , fu nc tio ns a v a ila b le , 1-19A u to MA T O R S e tu p , fu nc tio ns a v a ila b le , 1-19Cy to m e te r Co ntr o l, fu nc tio ns a v a ila b le , 1-19de s c r ip tio n, 1-18Mo dify Co lo r P r e c e de nc e , fu nc tio ns

a v a ila b le , 1-19p lo ts , fu nc tio ns a v a ila b le , 1-19P r o to c o l E x p lo r e r , fu nc tio ns a v a ila b le , 1-18r e s o u r c e e x p lo r e r , fu nc tio ns a v a ila b le , 1-18T ile S p e c ia l, fu nc tio ns a v a ila b le , 1-19

Ee dit

q u a dr a nt r e g io n, r e - p o s itio n, 1-25e dit p r o du c ts , 3-14e q u a tio ns

S ee fo r m u la se r r o r m e s s a g e s

CX P s o ftw a r e s c r e e n, 6-11Cy to m e te r S ta tu s W indo w , 6-12c y to m e te r .lo g , de s c r ip tio n, 6-13c y to m e te r a r c h iv e .lo g file , 6-14de s c r ip tio n, 6-11h o w to c le a r, 6-12lis t o f, 6-15r e c o rd in ins tr u m e nt lo g b o o k , 3-5ta b le , 6-15w h e r e dis p la y e d, 6-11

e th a no lu s e d in s h u tdo w n c le a ning , 2-11

E v e nts /S e c c o u nte r

PN 624923B IND E X -5

INDEX

u s e to m o nito r a c q u is itio n, 4-4, 4-8 , 4-11e x p o r t da ta

Q C da ta , 3-4

FF C 50 0

b a s ic o p e r a ting te c h niq u e s , 1-13c o ntr o ls a nd indic a to r s , 1-3do no t tr a ns fe r s e tting s file s , 3-9illu s tr a tio n, 1-3inte nde d u s e , 1-1m a nu a ls , h a rd-c o p y , x iiip u r p o s e , 1-1s p e c ific a tio ns , 4-1w a r m - u p tim e , 2-4, 3-5S ee also s y s te m

fie ld s to pa ng le s o f F S it c o lle c ts , 5-4fa c to r y s e tting , 5-4

filte r s48 8 D L, fu nc tio n, 5-650 0 LP , fu nc tio n, 5-7a ir , lo c a tio n, 2-6a ir , w h e n to c le a n, 6-11c o nfig u r a tio n, fo u r F L s e ns o r s , 5-7da ily c h e c k a t s ta r tu p , 2-6dic h r o ic , fu nc tio n, 5-7lo ng p a s s ( LP ) , fu nc tio n, 5-7s p a tia l, fu nc tio n, 5-6

F LS ee flu o r e s c e nt lig h t ( F L)

flo w c e llh o w to a v o id c lo g g ing , 4-2, 4-5, 4-9h y dr o dy na m ic fo c u s ing , 5-1illu s tr a tio n, 5-2

flo w r a tes a m p le , indic a to r s , 1-5

F lo w -Ch e c k 67 5 flu o r o s p h e r e sc h a r t fo r a p p lic a tio ns , 3-2Q C p r o c e du r e , 3-2Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

F lo w -Ch e c k 7 7 0 flu o r o s p h e r e sc h a r t fo r a p p lic a tio ns , 3-2Q C p r o c e du r e , 3-2Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

F lo w -Ch e c k flu o r o s p h e r e sc h a r t fo r a p p lic a tio ns , 3-2

Q C p r o c e du r e , 3-2Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

F lo w P A G Ec r e a ting , 1-31de le te , 1-32e x a m p le , 1-31

F lo w - S e t 67 5 flu o r o s p h e r e sc h a r t fo r a p p lic a tio ns , 3-3Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

F lo w - S e t 7 7 0 flu o r o s p h e r e sc h a r t fo r a p p lic a tio ns , 3-3Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

F lo w - S e t flu o r o s p h e r e sc h a r t fo r a p p lic a tio ns , 3-3Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

flu idic s s y s te m sc h e c k s ta b ility , 3-2v e r ify , 3-1

flu o r e s c e nc einte ns ity de te r m ine s p e a k p u ls e h e ig h t, 5-8

flu o r e s c e nt lig h t ( F L)c e ll illu m ina tio n, 5-4c o lle c tio n, 5-6indic a to r s , 1-5w h e n to u s e , e x a m p le , 5-4

flu o r o c h r o m ep r e p a r e c o m p e ns a tio n tu b e fo r e a c h , 3-3

flu o r o s p h e r e sp r e p a r e fo r A u to S e tu p , 3-2p r e p a r e p e r p a c k a g e ins e r ts , 3-2, 3-3, 3-6, 3-10r e -e s ta b lis h r a ng e s w ith ne w lo t, 3-1r u nning A u to S e tu p , 3-5r u nning F lo w -Ch e c k , 3-2u s a g e c h a r t, 3-2, 3-3S ee also indiv idu a l na m e s o f flu o r o s p h e r e s

fo c u s ing , h y dr o dy na m icde s c r ip tio n, 5-1

fo r m u la sCV , 5-16line a r r e g io n s ta tis tic s , 5-16m e a n, 5-16R A T IO p a r a m e te r, 5-13S D ( s ta nda rd de v ia tio n) , 5-16

fo r w a rd s c a tte r ( F S )c e ll illu m ina tio n, 5-4dis p la y indic a to r, 1-4


INDEX

lig h t c o lle c tio n, 5-4s e ns o r, 5-4

F SS ee fo r w a rd s c a tte r

Gg a in

a dju s t, 3-1r u n A u to S e tu p to s e t, 3-3

g a tingde s c r ip tio n, 5-15

g a u g e ss y s te m v a c u u m , 2-6

g e ne r a te d s ig na lsde s c r ip tio n, 5-10

g r a p h ic sfo r illu s tr a tio n o nly, x v

g r o u ndu s e r , if s ta tic c h a r g e e x is ts , 2-1

Hh a z a rds

r a dia tio n e x p o s u r e , 6-1He liu m N e o n la s e r

S ee He N e la s e rh e lp

a b o u t, 1-38a b o u t, s e le c t, 1-38a c c e s s ing , 1-37h o w to u s e , 1-37illu s tr a tio n o f la y o u t, 1-37u s ing th e o nline He lp s y s te m , 1-37

He lp Me nu , 1-38He N e la s e r

de s c r ip tio n o f h o w its lig h t is h a ndle d, 5-7illu s tr a tio n, 5-5la s e r la b e ls , 6-6, 6-7s h u tte r o p e n indic a to r, 1-4S ee also la s e r

h ig h v o lta g e sa dju s t, 3-1r u n A u to S e tu p to s e t, 3-3S ee also v o lta g e s

h is to g r a ms ta tis tic s , 5-16ty p e s o f dis p la y , 5-13S ee also p lo ts

h o m e p o s itio n

s a m p le c a r o u s e l, 1-15H ost C ommand Invalid, 6-15h o t k e y s

S ee k e y b o a rd s h o r tc u tsh y dr o dy na m ic fo c u s ing

de s c r ip tio n, 5-1

II2-o f-5

b a r -c o de , B -5ic o ns

c o ntr o l p a ne l, 1-10m y c o m p u te r, 1-10ta s k b a r, 1-8S ee also b u tto ns , T o o lb a r b u tto ns

idleCy to m e te r , p e r fo r m p r im e c y c le b e fo r e r u nning

s a m p le s , 4-1Im m u no -B r ite flu o r o s p h e r e s

Q C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

Im m u no -T r o l Ce llsQ C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

Im m u no -T r o l Lo w Ce llsQ C p r o c e s s u s e d fo r, 3-1S ee also c o ntr o ls

Incorrect C y tometer S erial N umbera p p e a r s w h e n tr a ns fe r ins tr u m e nt s e tting s , 3-9

indic a to r p a ne lsCy to m e te r, 1-4

indic a to r sCy to m e te r r e a dy, 1-4la s e r o n, 1-4le v e l s e ns e , 1-6, 6-20s a m p le flo w r a te , 1-5s h e a th flo w , 1-6s h e a th lo w , 1-6, 6-20s ig na l a m p litu de , 1-5w a s te fu ll, 1-6, 6-20

Initializ ationa p p e a r s du r ing s ta r tu p c y c le , 2-5

ins tr u c tio ns fo r u s e m a nu a linfo r m a tio n o r g a niz a tio n, x iiiu s ing th e m a nu a l, x iii

ins tr u m e ntdo no t tr a ns fe r s e tting s file s , 3-9o p tic a l s y s te m , 5-5

inte nde d u s e o f th e ins tr u m e nt, 1-1

PN 624923B IND E X -7

INDEX

Inte r le a v e d 2-o f-5b a r -c o de , B -5

Inte r le a v e d 2-o f-5 b a r c o dec h e c k s u m a lg o r ith m , B -7

intr o du c tio n to y o u r m a nu a ls , x iii

Kk e y b o a rd

s h o r tc u ts , 1-17u s e to s e le c t s o ftw a r e c o m m a nds , x v

Lla b e ls

b a r -c o de , s p e c ific a tio ns , B -6CD R H-r e q u ir e d, 6-1la s e r , w a rning , 6-1u niv e r s a l p o w e r s u p p ly , w a rning , 6-10S ee also b a r -c o de la b e ls

la b e ls , b a r -c o deS ee b a r - c o de la b e ls

la b o r a to r y ’s r a ng e sw h e n to r e -e s ta b lis h , 3-1

la s e rb a r -c o de r e a de r , MCL, B -6b e a m s h a p ing , 5-3b lu e , S ee A r g o n la s e rr e d, S ee s o lid-s ta te la s e r o r He N e la s e rr e - e s ta b lis h r a ng e s w h e n r e a lig ne d, 3-1r e -e s ta b lis h r a ng e s w h e n r e p la c e d, 3-1s a fe ty , 6-1w a rning la b e ls , 6-1

L aser C urrent E rror, 6-15L aser P ower E rror, 6-15L aser R egulation Warning, 6-15L aser S tart E rror, 6-15la y o u t

h e lp w indo w , 1-37le ns e s

b e a m s h a p ing , 5-3c r o s s - c y lindr ic a l, 5-3

le v e l s e ns e a nd flo w indic a to r s , 1-6le v e l s e ns e indic a to r s , 6-20

s h e a th lo w , 6-20w a s te fu ll, 6-20

lig h t c o lle c tio n, s e p a r a tio n a nd m e a s u r e m e ntde s c r ip tio n, 5-4

lig h t s c a tte rinte ns ity de te r m ine s p e a k p u ls e h e ig h t, 5-8

lig h tsS ee indic a to r s

line a r r e g io nc r e a ting , 1-26S ee also r e g io ns

line a r itym o nito r ins tr u m e nt, 3-1

lin-lo gm e a n m e th o d, 5-18

lis tm o dede s c r ip tio n, 5-15

lo g o nins tr u c tio ns , 2-4W indo w s s c r e e n, 2-4

lo g -lo gm e a n m e th o d, 5-17

lo ng p a s s ( LP ) filte r sfu nc tio n, 5-7

MMa inte na nc e Lo g

r e c o rd c le a ning , 2-11r e c o rd da ily s h u tdo w n, 2-11r e c o rd s ta r tu p c h e c k s , 2-8

m a nu a l m o dee dit s a m p le ID , 4-6e nte r tu b e lo c a tio n, 4-8h o w to s e le c t, 4-7r u nning s a m p le s , 4-5

m a nu a ls fo r y o u r ins tr u m e nta b o u t y o u r ins tr u c tio ns fo r u s e m a nu a l, x iiic o nv e ntio ns u s e d, x ivh o w to u s e , x iiiintr o du c tio n to y o u r m a nu a ls , x iiiu s ing th e m a nu a ls , x iii

MCL ( Mu lti-T u b e Ca r o u s e l Lo a de r )c h e c k v o r te x fu nc tio n, 2-8do no t o p e n w h ile m o v ing , 4-6illu s tr a tio n, 1-6la s e r la b e ls , 6-8 , 6-9lo a ding th e s a m p le , 5-1p u tting a c a r o u s e l in th e MCL, 1-14r e m o v ing a c a r o u s e l fr o m th e MCL, 1-16tu b e a c c e s s do o r, 4-9u s ing c a r o u s e ls , 1-13S ee also s a m p le c a r o u s e l a nd c a r o u s e l, s a m p le

MCL a u to m a tic m o deS ee a u to m a tic m o de

M C L C arousel In/O ut E rror, 6-16


INDEX

M C L C arousel L abel E rror, 6-16M C L C arousel R otate E rror, 6-16M C L C P U E rror, 6-16M C L D oor O pen E rror, 6-16M C L D oor O pen Warning, 6-16M C L E P R O M E rror, 6-16M C L E rror, 6-16M C L Interlock E rror, 6-16MCL m a nu a l m o de

S ee m a nu a l m o deM C L P arallel R ecv E rror, 6-16M C L P arallel S end E rror, 6-16M C L P robe U p/D own error, 6-16M C L P rogram L oad E rror, 6-16M C L R A M E rror, 6-16M C L R ec from F ault E rror, 6-16M C L R eceive Timeout E rror, 6-16M C L R equest E rror, 6-16MCL s a m p le h e a d

w h e n to c le a n, 6-11M C L S erial 1 R ecv E rror, 6-16M C L S erial 1 S end E rror, 6-16M C L S erial 2 R ecv E rror, 6-17M C L S erial 2 S end E rror, 6-17M C L Transmit Timeout E rror, 6-17M C L Tube D isplaced E rror, 6-17M C L Tube Jam E rror, 6-17M C L Tube L oad E rror, 6-17M C L Tube P osition E rror, 6-17M C L Tube U p/D own E rror, 6-17m e a n

fo r m u la , 5-16lin-lo g m e th o d, 5-18lo g -lo g m e th o d, 5-17

m e a n p o s itio na s c e r ta in ta r g e t, 3-1

m e s s a g e se r r o r , de s c r ip tio n, 6-11e r r o r , lo g , 6-13e r r o r , w h e r e dis p la y e d, 6-11

m is m a tc h e d ID s , 4-4, 4-8 , 4-11m o de

a u to m a tic , 4-2de finitio n, 5-16m a nu a l, 4-5s ing le tu b e m o de , 4-9

Mo dify Co lo r P r e c e de nc edr a g a nd dr o p fu nc tio ns , 1-19

m o nito r

c o lo r r e s o lu tio n, 1-11fr o nt v ie w , 1-7ins tr u m e nt line a r ity , 3-1

m o u s eu s ing , 1-10

m u ltip le line a r r e g io nsc r e a ting , 1-27

My Co m p u te rW indo w s , 1-10

Nna v ig a tio n p a ne

illu s tr a tio n, 1-37lo c a tio n, 1-37

ne g a tiv e c o ntr o la dditio na l s te p s in A u to S e tu p w iz a rd, 3-7r u nning , 3-7

ne tw o r klo g in a t s ta r tu p , 2-4s e a r c h fo r file s , fo lde r s , a nd c o m p u te r s , 1-9

ne w p a ne lc a r r y r e g io ns fr o m p r e v io u s p r o to c o l, 1-34u s e ins tr u m e nt s e tting s , 1-34u s e ins tr u m e nt s e tting s fr o m e x te rna l file , 1-34u s e ins tr u m e nt s e tting s fr o m p r e v io u s te s t, 1-34u s e p lo ts a nd g a te s fr o m p r o to c o l, 1-34u s e r e g io ns fr o m p r o to c o l, 1-34u s e r e g io ns fr o m th is p r o to c o l, 1-34

ne w p a ne l w iz a rdS ee p a ne l w iz a rd

N W 7b a r -c o de , B -5

Oo nline He lp

S ee h e lpo p e r a tio n

p r inc ip le s o f, 5-1o p tic a l

s y s te m , 5-5s y s te m s , c h e c k s ta b ility , 3-2

o p tic a l a r e ala s e r la b e ls , 6-3, 6-4

o p tic a l c h a r a c te r is tic sb a r -c o de la b e ls , B -6fo r b a r - c o de la b e ls , B -6

o p tic a l filte r p la tede s c r ip tio n, 5-7

PN 624923B IND E X -9

INDEX

filte r c o nfig u r a tio n, 5-7

Pp a c k a g e ins e r ts

c o nta in flu o r o s p h e r e s ins tr u c tio ns , 3-2, 3-3r e a g e nt, u s e to p r e p a r e s a m p le s , 4-2, 4-5, 4-9u s e to e s ta b lis h e x p e c te d r a ng e s , 3-6u s e to e s ta b lis h no r m a l r a ng e s , 3-1u s e to e s ta b lis h ta r g e t c h a nne ls , 3-6, 3-10u s e to p r e p a r e CY T O -T R O L c e lls , 3-3u s e to p r e p a r e I m m u no -T r o l c e lls , 3-3

p a ne l r e p o r tg e ne r a te , 1-32

p a ne l w iz a rdc r e a te ne w p a ne ls , 1-32

p a ne lsc a n s k ip tu b e s b e tw e e n p a ne ls , 1-14c le a ns e p a ne l, de s c r ip tio n, 5-11c r e a ting , 1-32do no t s k ip tu b e s w ith in p a ne l, 1-14h o w to s e le c t, 4-4, 4-7 , 4-10ne w , 1-32p a ne l w iz a rd, 1-32Q C, ne e d u niq u e p r o to c o ls , 3-13s p e c ia l, 5-11w h a t to do if th e r e is a de le te d

p r o to c o l, 4-4, 4-7 , 4-10S ee also ne w p a ne l

p a r a m e te r sA U X , 5-12de s c r ip tio n, 5-12na m e s , 1-34R A T IO , 5-13r u n A u to S e tu p to s e t g a ins , 3-3T IME , 5-12

p a s s w o rdse nte r a t lo g o n, 2-4fo r a u to s ta r tu p /s h u tdo w n, 2-14

P C7S ee also F lo w -Ch e c k 7 7 0 a nd F lo w -S e t 7 7 0

flu o r o s p h e r e sp e a k s ig na l

de s c r ip tio n, 5-8S ee also s ig na ls

p e r fo r m a nc e c h a r a c te r is tic s , 1-38p h e no ty p e

de finitio n, 5-13p h o to s e ns o r s

S ee s e ns o r s

p ic k u p le ns /s p a c ia l filte r a s s e m b lyfu nc tio n, 5-6

p lo tsdr a g a nd dr o p fu nc tio n, 1-19P R IS M, 5-13P R IS M, illu s tr a tio n, 5-14ty p e s o f dis p la y , 5-13S ee also h is to g r a m s , c o lo r do t p lo ts , c o nto u r

p lo ts , de ns ity p lo ts , do t p lo ts , h is to g r a m p lo ts a nd p r is m p lo ts

P MTr e - e s ta b lis h r a ng e s w h e n r e p la c e d, 3-1S ee also h ig h v o lta g e s

p o ly g o na l r e g io nc o nv e r t to a n A u to G a te r e g io n, 1-30c r e a ting , 1-22S ee also r e g io ns

p o s itio n 10u s e d fo r s ing le tu b e m o de , 4-10

p o w e r fa ilu r eh o w to h a ndle s a m p le s , 4-2

P o w e r S u p p lyfr o nt v ie w , 1-7u niv e r s a l, 1-7u niv e r s a l, w a rning la b e ls , 6-10v o lta g e - s p e c ific , 1-7

p r e c a u tio nsa g a ins t r a dia tio n, 6-1

P r e p a r ing A u to S e tu p R e a g e nts , 3-2P reparing S ample

a p p e a r s du r ing s a m p le c y c le , 4-4, 4-8 , 4-11p r im e c y c le

inc lu de d in s ta r tu p c y c le , 2-5w h e n to r e q u e s t, 4-1

p r inc ip le s o f o p e r a tio n, 5-1P r inte r

da ily c h e c k s , 2-8o p e r a ting ins tr u c tio ns , 1-7

P R IS Mde s c r ip tio n, 5-13p lo t, 5-14s ig na ls th a t c a n b e a s s ig ne d to it, 5-14w h e n to u s e , 5-13

P r o g r a m sW indo w s , 1-9

P r o to c o l E x p lo r e rdr a g a nd dr o p fu nc tio ns , 1-18

p r o to c o lsA S , de s c r ip tio n, 5-11c le a ns e b le a c h .p r o , 5-11

PN 624923BIND E X -1 0

INDEX

c le a ns e di w a te r .p r o , 5-11Cle a ns e , de s c r ip tio n, 5-11c r e a ting , 1-20de finitio n, 5-11de s c r ip tio n, 5-11dr a g a nd dr o p fu nc tio n, 1-18h o w to s e le c t, 4-4, 4-7 , 4-10info r m a tio n c o nta ine d, 5-11ne w , 1-20Q C, de s c r ip tio n, 5-11s p e c ia l, 5-11u s e d in A u to S e tu p p a ne ls , 5-11w h a t to do if th e r e is a de le te d p r o to c o l in a

p a ne l, 4-4, 4-7 , 4-10S ee also ne w p r o to c o l

P u b lis h D a ta to MS E x c e lu s e fo r Le v e y Je nning s p lo ts , 5-11

p u ls eS ee s ig na ls

QQ C m a te r ia ls

Q C p r o c e s s e s u s e d fo r, 3-1S ee also q u a lity c o ntr o l ( Q C)

q u a dr a nt r e g io nc r e a ting , 1-24e diting , r e - p o s itio n, 1-25S ee also r e g io ns

q u a lity c o ntr o l ( Q C) , 3-1a b o u t, 3-1a s s ig ning Q C p r o du c ts , 3-14a u to s e tu p s c h e du le r, 3-4a u to s e tu p w iz a rdc h e c k s p e c ific a p p lic a tio ns , 3-1da ily p r o c e du r e s , 3-2e dit p r o du c ts , 3-14e x p o r t Q C da ta to s p r e a ds h e e t, 3-4F lo w -Ch e c k flu o r o s p h e r e s , 3-2F lo w - S e t flu o r o s p h e r e s , 3-3p r o du c ts , 3-14Q C m a te r ia ls u s e d fo r Q C p r o c e s s , 3-1Q C p r o c e s s e s , 3-1r e v ie w r e s u lts , 3-15s e tting u p Q C p r o du c ts , 3-14

Rr a dia tio n h a z a rds , 6-1r a ng e s , la b o r a to r y ’s

e s ta b lis h e x p e c te d r a ng e s , 3-6w h e n to r e -e s ta b lis h , 3-1

R A T IO p a r a m e te rde s c r ip tio n, 5-13fo r m u la , 5-13

r e a g e ntsc h e c k le v e l a t s ta r tu p , 2-3

r e c o rdc le a ning , 2-11da ily s h u tdo w n, 2-11s ta r tu p c h e c k s , 2-8

r e c ta ng u la r r e g io nc r e a ting , 1-23S ee also r e g io ns a nd q u a dr a nt r e g io n

R e c y c le B inW indo w s , 1-9

r e d la s e rS ee s o lid-s ta te la s e r o r He N e la s e r

r e g io nsA u to G a te , 1-28c a r r y fr o m p r e v io u s p r o to c o l, 1-34c r e a ting

A u to G a te r e g io n, 1-28line a r r e g io n, 1-26m u ltip le line a r r e g io ns , 1-27p o ly g o na l r e g io n, 1-22q u a dr a nt r e g io n, 1-24r e c ta ng u la r r e g io n, 1-23

de s c r ip tio n, 5-15line a r r e g io n, 1-26m u ltip le line a r r e g io ns , 1-27p o ly g o na l r e g io n, 1-22q u a dr a nt r e g io n, 1-24

r e -p o s itio n, 1-25r e c ta ng u la r r e g io n, 1-23ty p e s , 5-15ty p e s o f fu nc tio ns , 5-15

r e p la c e m e nt p r o c e du r e ss c h e du le , 6-11

r e p o r ting u nitss h o w n in U S u nit fo r m a t, x v

r e q u ir e m e nts , s a m p le , 4-1r e s o lu tio n

s e t c o lo r m o nito r, 1-11r e s o u r c e e x p lo r e r

dr a g a nd dr o p fu nc tio ns , 1-18r e s u lts

r e v ie w Q C r e s u lts , 3-15R o m lo c k

PN 624923B IND E X -1 1

INDEX

nu m b e r o n R o m lo c k , 1-38R u n

W indo w s , 1-9r u nning

a p p lic a tio n de finitio n w iz a rd, 3-10a u to s e tu p w iz a rdF lo w -Ch e c k flu o r o s p h e r e s , 3-2F lo w - S e t flu o r o s p h e r e s , 3-3s a m p le s in MCL a u to m a tic m o de , 4-2s a m p le s in MCL m a nu a l m o de , 4-5s a m p le s in s ing le tu b e m o de , 4-9

Ss a fe ty p r e c a u tio ns

la s e r, 6-1s a m p le

c h e c k s b e fo r e r u nning , 4-1flo w , de s c r ip tio n, 5-1h o w to h a ndle a fte r p o w e r fa ilu r e , 4-2illu m ina te d in th e flo w c e ll, 5-4lo a ding , a u to m a te d, 5-1m e a s u r a b le c e ll s iz e s , 4-1o p tim u m c o nc e ntr a tio n, 4-1p r e p a r a tio n, 4-1p r e p a r e p e r r e a g e nt p a c k a g e ins e r t, 4-2, 4-5, 4-9r e q u ir e m e nts , 4-1r is k o f m is ide ntific a tio n, 4-2r u n s a m p le s in MCL a u to m a tic m o de , 4-2r u n s a m p le s in MCL m a nu a l m o de , 4-5r u n s a m p le s in s ing le tu b e m o de , 4-9v e r ify m a nu a lly e nte r e d info r m a tio n b e fo r e

r e p o r ting r e s u lts , 4-5, 4-9, 4-11s a m p le c a r o u s e l

b a r -c o de la b e ls , 5-1de s c r ip tio n, 5-1e nte r th e ID nu m b e r, 4-4, 4-7h o m e p o s itio n, 1-15p u tting a c a r o u s e l in th e MCL, 1-14r e m o v ing a c a r o u s e l fr o m th e MCL, 1-16S ee also c a r o u s e l, s a m p le a nd MCL

s a m p le flo wde s c r ip tio n, 5-1r a te indic a to r s , 1-5

s a m p le IDc a n e dit a fte r e a c h tu b e in m a nu a l m o de , 4-5c a n e dit a fte r e a c h tu b e in s ing le tu b e m o de , 4-9s to p e a c h tu b e s o y o u c a n e dit in m a nu a l

m o de , 4-6s to p e a c h tu b e s o y o u c a n e dit in s ing le tu b e

m o de , 4-9S ample P ressure E rror, 6-17s a m p le p r o b e

h o w to a v o id c lo g g ing , 4-2, 4-5, 4-9w h e n to c le a n, 6-11

s a m p le tu b eS ee te s t tu b e

s a m p le tu b ingh o w to a v o id c lo g g ing , 4-2, 4-5, 4-9

s a m p ling s y s te mw h e n to c le a n, 6-11

s c h e du lea dju s tm e nt p r o c e du r e s , 6-11a u to s h u tdo w n, 2-15a u to s ta r tu p , 2-15c le a ning , 6-11r e p la c e m e nt, 6-11

S D ( s ta nda rd de v ia tio n)fo r m u la , 5-16

S e a r c hW indo w s , 1-9

s e c u r ityW indo w s 20 0 0 p a s s w o rd, 1-8

s e ns ing c o m p a r tm e ntla s e r la b e ls , 6-2

s e ns o r sCy to m e te r, 5-8

S ensors S tream Warning, 6-17s e tting s

do no t tr a ns fe r file s b e tw e e n ins tr u m e nts , 3-9s a v e fr o m A u to S e tu p I I, 3-9W indo w s , 1-9

S heath D rawer E rror, 6-17s h e a th flu id

flo w indic a to r, 1-6lo w indic a to r, 1-6, 6-20

s h e a th flu id c o nta ine rw h e n to c le a n, 6-11

s h e a th flu id flo w indic a to rlo c a tio n, 1-6

S heath L evel E rror, 6-17S heath L evel Warning, 6-17s h e a th lo w indic a to r

de s c r ip tio n, 6-20lo c a tio n, 1-6, 6-20

S heath P ressure E rror, 6-18s h o r tc u ts , s o ftw a r e , 1-17

dr a g a nd dr o p , 1-18k e y b o a rd, 1-17

S h u t D o w n

PN 624923BIND E X -1 2

INDEX

W indo w s , 1-9s h u tdo w n

da ily s h u tdo w n, 2-9e x te nde d, p r o c e du r e , 2-12h o w to s e t u p a u to s h u tdo w n, 2-14m inim u m tim e , 2-9w h e n to s h u t do w n th e Cy to m e te r, 2-9

s ide s c a tte r ( S S )c e ll illu m ina tio n, 5-4c o lle c tio n, 5-6s e ns o r, 5-6w h e n to u s e , e x a m p le , 5-4

s ig na l a m p litu de indic a to r s , 1-5s ig na ls

a m p lific a tio n, 5-10g e ne r a te d, 5-10inte g r a l, de s c r ip tio n, 5-10inte g r a l, illu s tr a tio n, 5-10inte g r a l, ty p e s , 5-10p e a k , de s c r ip tio n, 5-8p e a k , illu s tr a tio n, 5-9, 5-10p r o c e s s ing o f, o p e r a tio n p r inc ip le s , 5-8v o lta g e p u ls e , 5-8

s ig ning o n to CX P s o ftw a r e , 2-5s ing le tu b e m o de

e dit s a m p le I D , 4-9h o w to r e q u e s t, 4-10r u nning s a m p le s , 4-9u s e s p o s itio n 10 in c a r o u s e l, 4-10

s k ip p o s itio nsb e tw e e n p a ne ls , 1-14no t w ith in a p a ne l, 1-14

s o ftw a r eo nline h e lp s y s te m , 1-37s h o r tc u ts , 1-17s ig ning o n to CX P s o ftw a r e , 2-5

s o lid-s ta te la s e rillu s tr a tio n, 5-5la s e r la b e ls , 6-5, 6-6S ee also la s e r

s p a tia l filte rS ee filte r s

s p e c ia l p a ne ls , 5-11s p e c ia l p r o to c o ls , 5-11s p e c ific a tio ns

b a r -c o de la b e ls , B -2, B -6s p e c im e n

S ee s a m p les p r e a ds h e e t

e x p o r t Q C da ta , 3-4S S

S ee s ide s c a tte rs ta b ility

c h e c k flu idic s s y s te m s , 3-2c h e c k o p tic a l s y s te m s , 3-2

s ta insc o m p e ns a tio n tu b e s , 3-3v e r ific a tio n tu b e , 3-3

S ta r t b u tto nde s c r ip tio n, 1-8m e nu , illu s tr a tio n, 1-8W indo w s , 1-8

s ta r te r k itMCL s u p p lie s , 1-13

s ta r tu pb e fo r e y o u b e g in, 2-1da ily p r o c e du r e , 2-1da ily s ta r tu p , 2-1h o w to s e t u p a u to s ta r tu p , 2-14lo g into ne tw o r k , 2-4r e c o rd da ily c h e c k s , 2-8

S tartup in P rocessa p p e a r s du r ing s ta r tu p c y c le , 2-5

S tate M achine F ailure, 6-18s ta tic c h a r g e

u s e r ne e ds to b e g r o u nde d, 2-1s ta tis tic s

h is to g r a m , 5-16line a r r e g io n, 5-16lo g r e g io n, 5-17

s ta tu s m e s s a g e sr u nning flu o r o s p h e r e s , 3-5s a m p le c y c le m e s s a g e s , 4-4, 4-8 , 4-11s ta r tu p c y c le m e s s a g e s , 2-5

s to p c o u nte ffe c t o n A u to G a te r e g io n, 1-28

S toppinga p p e a r s du r ing s a m p le c y c le , 4-4, 4-8 , 4-11

s y s te mc o ntr o ls a nd indic a to r s , 1-3o p tic a l, 5-5S ee also F C 50 0

s y s te m p r e s s u r ec o r r e c t r a ng e , 2-6g a u g e lo c a tio n, 2-6h o w to a dju s t, 2-6

S y stem P ressure E rror, 6-18s y s te m v a c u u m

PN 624923B IND E X -1 3

INDEX

da ily c h e c k , 2-6g a u g e lo c a tio n, 2-6

S y stem V acuum E rror, 6-18s y s te m v a c u u m g a u g e

w h e n to c a ll s e r v ic e , 2-6

Tta r g e t c h a nne ls , la b o r a to r y ’s

e s ta b lis h ta r g e t c h a nne ls , 3-6, 3-10T a s k b a r

b u tto ns , 1-8de s c r ip tio n, 1-8ic o ns , 1-8illu s tr a tio n, 1-8S ta r t b u tto n, 1-8W indo w s , 1-8

Temperature E rror, 6-18Temperature S ensor F ailure, 6-18Temperature Warning, 6-18te s t tu b e

h o w to a p p ly b a r - c o de la b e l, 1-13ID m is m a tc h , h o w th e s y s te m

h a ndle s , 4-4, 4-8 , 4-11illu s tr a tio n in s a m p le c a r o u s e l, 1-14lo a ding c a r o u s e l, 1-14p u t into th e c a r o u s e l, 1-14s iz e , 4-1

T ile S p e c ia ldr a g a nd dr o p fu nc tio ns , 1-19

T IME p a r a m e te rde s c r ip tio n, 5-12

to o lb a rb u tto ns , 1-19T a s k b a r b u tto ns , 1-8

to p ic p a neillu s tr a tio n, 1-37lo c a tio n, 1-37

Transputer 1 E rror, 6-18Transputer 2 E rror, 6-18Transputer L ink E rror, 6-19Transputer L ink Warning, 6-19tr o u b le s h o o ting , 6-1

e r r o r m e s s a g e lo g , 6-13e r r o r m e s s a g e s , 6-11le v e l s e ns e indic a to r s , 6-20

tu b e a c c e s s do o rillu s tr a tio n, 4-10lo c a tio n, 4-10u s e fo r s ing le tu b e m o de , 4-9

tu b e , s a m p leS ee te s t tu b e

UU nk nown S tream Warning, 6-19U S D -1

b a r -c o de , B -5U S D -1.25

b a r -c o de , B -5U S D -3

b a r -c o de , B -5U S D -4

b a r -c o de , B -5U S D -6

b a r -c o de , B -5u s e

ins tr u m e nt s e tting s , 1-34ins tr u m e nt s e tting s fr o m e x te rna l file , 1-34ins tr u m e nt s e tting s fr o m p r e v io u s te s t, 1-34p lo ts a nd g a te s fr o m p r e v io u s te s t, 1-34p lo ts a nd g a te s fr o m p r o to c o l, 1-34r e g io ns fr o m p r o to c o l, 1-34r e g io ns fr o m th is p r o to c o l, 1-34

u s e a nd fu nc tio n, ins tr u m e nt, 1-1U s e r I D , 2-5u s e r na m e

S ee U s e r I Du s ing

m o u s e , 1-10th e o nline h e lp s y s te m , 1-37

VV acuum C hamber Warning, 6-19v a c u u m filte r

da ily c h e c k , 2-6lo c a tio n, 2-6w h e n to c a ll s e r v ic e , 2-6

v a c u u m g a u g eda ily c h e c k , 2-6

v a c u u m tr a pda ily c h e c k , 2-8lo c a tio n, 2-8w h e n to c le a n, 6-11

v a c u u m , c h e c k ingda ily s ta r tu p , 2-6s y s te m v a c u u m g a u g e , 2-6v a c u u m filte r, 2-6v a c u u m tr a p , 2-8

PN 624923BIND E X -1 4

INDEX

V erificationa p p e a r s du r ing s ta r tu p c y c le , 2-5

v e r ific a tio n tu b e sA u to S e tu p , 3-3c o m p e ns a tio n c o e ffic ie nts , 3-8

v e r ifya lig nm e nt, 3-1a p p lic a tio n c o ntr o l, 3-1c o r r e c t s e tting s , 3-1flu idic s s y s te m s , 3-1HP CV v e r s u s e x p e c te d v a lu e , 3-1

v ide o c a rdss e tting ne e de d fo r 3D a c c e le r a tio n, 1-12

v ir u s p r o te c tio n p r o g r a ms h u tdo w n c o m p u te r, 2-1

v ir u s s c a nno t w h ile r u nning CX P , 2-1

v o lta g e p u ls e s ig na lsde s c r ip tio n, 5-8S ee also s ig na ls

v o lta g e sa dju s t, 3-1a dju s t to u s e ne g a tiv e c o ntr o l, 3-7r u n A u to S e tu p to s e t, 3-3S ee also h ig h v o lta g e s

Ww a r m - u p

ins tr u m e nt, 2-4, 3-5Waste B ack pressure E rror, 6-19w a s te c o nta ine r

e m p ty a t s ta r tu p , 2-2w a s te fu ll indic a to r, 6-20

w a s te fu ll indic a to rde s c r ip tio n, 6-20lo c a tio n, 1-6, 6-20

Waste L evel E rror, 6-19Waste L evel Warning, 6-19w a te r tr a p

da ily c h e c k , 2-6lo c a tio n, 2-6w h e n to c a ll s e r v ic e , 2-6

w e b s itea c c e s s to A p p lie d Cy to m e tr y S y s te m s , 1-38a c c e s s to B e c k m a n Co u lte r, 1-38

w h e n to c a ll s e r v ic ea ir filte r, 2-6s y s te m v a c u u m g a u g e , 2-6v a c u u m filte r, 2-6

w a te r tr a p , 2-6w h e n to s h u tdo w n, 2-9w h o le b lo o d

r u n a s a c o ntr o l, 3-1W indo w s

Co ntr o l P a ne l, 1-10D e s k to p , 1-8D o c u m e nts , 1-9He lp , 1-9My Co m p u te r, 1-10p a s s w o rd s e c u r ity , 1-8P r o g r a m s , 1-9R e c y c le B in, 1-9R u n, 1-9S e a r c h , 1-9S e tting s , 1-9S h u t D o w n, 1-9S ta r t b u tto n, 1-8T a s k b a r, 1-8

w iz a rdA u to S e tu p , r u nning , 3-5p a ne l S ee p a ne l w iz a rd

w o r k lis th o w to s e le c t, 4-4, 4-7 , 4-10p a ne l, 1-35te s t, 1-36

W o r k s ta tio nfr o nt v ie w , 1-7

PN 624923B

TRADEMARKS

A LT R A , B E CK MA N CO U LT E R lo g o , CO U LT E R , CO U LT E R CLE N Z , CY T O -CO MP, CY T O -T R O L, E lite , E P ICS , F lo w Ce ntr e , F lo w -Ch e c k , F lo w -Co u nt, F lo w - S e t, Im m u no -B r ite , Im m u no P r e p , I m m u no -T r o l, Is o F lo w , Mu lti-Q - P r e p , Q -P r e p , S te m -Co u nt, S te m -T r o l, T Q -P r e p , X L, a nd X L-MCL a r e tr a de m a r k s o f B e c k m a n Co u lte r, Inc .

A ll o th e r tr a de m a r k s , s e r v ic e m a r k s , p r o du c ts , o r s e r v ic e s a r e tr a de m a r k s o r r e g is te r e d tr a de m a r k s o f th e ir r e s p e c tiv e h o lde r s .

Printed on R ecy cled Pap er

Cy tom ics FC 50 0 w ith CX P Softw a re Docum enta tion

s R e fe r e nc eP N 17 557 0

U s e a nd F u nc tio n • Ins ta lla tio n • S p e c ific a tio ns • Lo g S h e e ts • U s ing CX P S o ftw a r e • G lo s s a r y • Inde x

s S p e c ia l P r o c e du r e sP N 17 557 2

Cle a ning P r o c e du r e s • R e p la c e /A dju s t P r o c e du r e s • Inde x

s Ins tr u c tio ns F o r U s eP N 624923

S y s te m O v e r v ie w • D a ily R o u tine • Q u a lity Co ntr o l • R u nning S a m p le s • O p e r a tio n P r inc ip le s • T r o u b le s h o o ting • Ho w to ... • B a r-Co de S p e c ific a tio ns • R e fe r e nc e s • Inde x

s Ma s te r Inde xP N 17 7 653

Co m b ine d inde x fo r th e Ins tr u c tio ns F o r U s e , S p e c ia l P r o c e du r e s , a nd R e fe r e nc e m a nu a ls .

s G e tting S ta r te dP N 17 7 652

O v e r v ie w o f th e s y s te m h a rdw a r e a nd s o ftw a r e .

s O p e r a to r Ma nu a lsCD -R O MP N 17 7 628

HT ML file o f th e S y s te m He lp • P D F file s o f th e R e fe r e nc e m a nu a l, Ins tr u c tio ns fo r U s e , a nd S p e c ia l P r o c e du r e s m a nu a l.

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