FlowJo 中文实用手册
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I FlowJo中文实用手册 杭州艾米绿生物科技有限公司 [email protected] 400-680-5527 FlowJo中文实用手册 （软件版本：FlowJo7.6.5 2012/06/20 更新） 第一章 FlowJo简介 ...................................................................................................................................... 1 一、FlowJo对电脑配置的要求 ............................................................................................................... 1 二、FlowJo的安装过程(Windows) ......................................................................................................... 1 三、FlowJo的工作台 ............................................................................................................................... 5 第二章 FlowJo分析单标样本 ...................................................................................................................... 6 一、由原始数据生成单参数直方图的过程 ........................................................................................... 6 二、图形的输出和保存 ......................................................................................................................... 10 第三章 FlowJo分析多色标记样本 ............................................................................................................ 13 一、由原始数据生成二维点图的过程 ................................................................................................. 13 二、二维点图的设门原则 ..................................................................................................................... 16 三、多色标记分析说明 ......................................................................................................................... 16 第四章 FlowJo软件荧光补偿 .................................................................................................................... 18 一、用FlowJo做荧光补偿并进行数据分析的步骤.............................................................................. 18 二、FlowJo荧光补偿的具体步骤 ......................................................................................................... 18 三、双指数转换 ..................................................................................................................................... 24 第五章 FlowJo的批处理功能 .................................................................................................................... 27 一、设门分析的批处理 ......................................................................................................................... 27 二、表格分析的批处理 ......................................................................................................................... 32 三、图形分析的批处理 ......................................................................................................................... 35 第六章 FlowJo分析细胞凋亡样本 ............................................................................................................ 43 一、细胞凋亡概述 ................................................................................................................................. 43 二、FlowJo分析凋亡数据的过程 ......................................................................................................... 44 三、凋亡数据结果分析 ......................................................................................................................... 45
description
the chinese version of Flowjo guide, simple but straight forward, explaning the basic of how to use Flowjo, like gating and backgating, worksheet, table and compensation etc.
Transcript of FlowJo 中文实用手册
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I FlowJo [email protected] 400-680-5527
FlowJo FlowJo7.6.5 2012/06/20
FlowJo ......................................................................................................................................1 FlowJo ...............................................................................................................1 FlowJo(Windows) .........................................................................................................1 FlowJo ...............................................................................................................................5
FlowJo ......................................................................................................................6 ...........................................................................................6 .........................................................................................................................10
FlowJo ............................................................................................................13 .................................................................................................13 .....................................................................................................................16 .........................................................................................................................16
FlowJo ....................................................................................................................18 FlowJo..............................................................................18 FlowJo .........................................................................................................18 .....................................................................................................................................24
FlowJo ....................................................................................................................27 .........................................................................................................................27 .........................................................................................................................32 .........................................................................................................................35
FlowJo ............................................................................................................43 .................................................................................................................................43 FlowJo .........................................................................................................44 .........................................................................................................................45
PatrickUnderline
PatrickUnderline
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PatrickUnderline
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FlowJo ....................................................................................................47 .........................................................................................................................47 FlowJo .................................................................................................48 .................................................................................................51 .........................................................................................................................53
PatrickUnderline
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FlowJo
FlowJoLeonard HerzenbergFACS
90FlowJo
FlowJo
FlowJo
1. PC
Windows XPVistaWindows7
512MB
2.
OSX10.3
512MB
FlowJo(Windows)
http://www.flowjochina.com/content/download
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V7.6.5
http://www.flowjochina.com/sites/default/files/7.6.5_X32.exe
FlowJo
FlowJo
NEXT
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NEXT
NEXT
Install
Done
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a. DongleFlowJo
b. Serial NumberUse
Serial Number
c. Get Trial LicenseFlowJo
Use Serial Number
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d. DemoFlowJo
Run in Demo ModeDemoFlowJoDemo
FlowJo
FlowJo
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FlowJo
,flowjochina
www.flowjochina.com/sites/default/files/demodata.rar
3 color compCy5PE comp.fcs
1. FlowJo
2. 3 color compFlowJo
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3 color comp3 color
comp
Cy5PE comp.fcs
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1) XSSC
SSC
2) YFSC
FSC
3)
:
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1) XCy5PE:CD4YHistogram
2)
a.
CD4CD4
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b.
CD4CD
a.
WORDPPT
b.
SVG
PNGEMFJPG
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c. FlowJo
d.
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e.
data001.JPGJPG, PNG, GIF, EMF,
SVG, PDF
FlowJoPNG
f.
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FlowJo
FlowJo
FlowJo3-color-experiment931115-
B02-Sample01.FCS
1. 3-color-experimentFlowJo
2. 931115-B02-Sample01.FCS
XFSCYSSC
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3. XFluor::CD3YFSC
CD3T
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4. TXCy5PE::CD4Y
PhyEry::CD8
Q1CD8CD8+ T
Q2
Q3CD4CD4+ T
Q4
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TREGCD4/CD25/FOXP3SSC vs. CD4
CD4+CD25 vs. FOXP3
CD25+/FOXP3+CD4+/CD25+/FOXP3+
TREG
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FlowJo
FlowJo
FlowJo
1.
2.
3.
FlowJo
FlowJoFlowJo
4
http://www.video.flowjochina.com/v75%20Compensation_conv.html
1. FlowJo
3 color compCy5PE comp.fcs, FITC
comp.fcs, PE comp.fcs3-COLOR.FCS
Cy5PE, FITC, PE
PatrickHighlight
PatrickHighlight
PatrickHighlight
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2.
Sample
FlourFITC comp.fcs
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3. 23FlowJo
4.
>
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(poor control )FlowJo
1. 1,2,3
FlowJoPE
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1) PEcw_Size
2) Clear
2.
1) PEFSC/SSC
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2) XPhyEry::aCD8Y
Histogram PEPE
a.
b.
3) PE-PE
PE+
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3. FlowJo
3-color comp
>
0Log
0
LogicalFlowJo
3-color comp3-
COLOR.FCS
1. 3-COLOR
XCD3YCD8
gate135.1%
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2. XYT
3. T
Change transform values
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X-10-30
a.
b. FCS3.0FlowJoFCS2.0
FlowJo
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FlowJo
Titration DataCD8Sample 1
Sample 2 ~ Sample 82
1.
Sample 1
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XFL1-HYHistogram Sample 1
FL1-H+
RenameCD8+FL1-H-
CD8-
2.
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CD8+
MedianParameter
FL1-HCD8FL1-H
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(FL1-H):MedianCD8-CD8
FL1-H
3.
Sample 1Sample 2
Sample 2Sample
1
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Sample 1
Titration Data
Sample 1
ALTSample 1
Titration Data
Sample 1
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1.
Sample 1CD8+CD8-
(FL1-H):Median
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2.
Keyword
Antibody Concentration
OK
3.
CD8+ (FL1-H) Median CD8- (FL1-H) MedianCD8
CD8MFI
OK
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4.
MFI
1MFI1.
1.
Sample 1
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FlowJoTitration Data
FlowJo
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2. (Overlay)
1
Titration Data
(1) Sample 1Sample 2
Sample 1
(2) SpecifyControl
Sample 1Sample 1OK
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(3)
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2
Titration Data
(1) Sample 1CTRL
Sample 1
Sample 1
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(2)
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(3)
(4) coloringfilled
(5)
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FlowJo
Annexin V-FITC/PI
Annexin V-FITC/PI
PSVAnnexin V-FITC
PS
PSPI
PS
AnnexinPI
Annexin V-FITC/PI
AnnexinAnnexin, PI
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FlowJo
apoptosisZL-2011-12-20_400.fcs
1. FlowJo
XFSC-A,YSSC-A
2. SSC/FSC
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XFITC,AnnexinV-FITCYPE-Texas,PI
1.
2.
Q1(AnnexinV-FITC)-/PI+
Q2: (AnnexinV+FITC)+/PI+
Q3(AnnexinV-FITC)+/PI-
Q4(AnnexinV-FITC)-/PI-
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Q2+Q3
AnnexinV
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FlowJo
G1
SG2MG1S
DNAG2DNA
M
DNAPIPI
DNARNA
DNA
DNA
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FlowJo
cell cycle 5206.001PI
1.
5206.001XFSC-HY
SSC-H
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2.
XFL2-W(width)YFL2-A(area)
BDFL2-A
FL2-WFL2-AFL2H
BeckmanXFL3-LIN,PI
YFL3-PEAK,PI
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3. FlowJo
G1SG2G1CV
G2CV
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http://www.flowjochina.com/content/tutorial-video#cellcycle
1.
RMS
RMS(root mean square error)
RMS3.85
2.
WatsonDean-Jett-Fox
RMSRMS
Could not fit.
Watson
Dean-Jett-Fox
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3.
G1G2meanCV
1 G2G12
G1G2G1
2(2G1)G2G1G20.5
2 G1G2
G1mean
G1mean
3 CV
DNAG1G2CV
G1 CV
=G2 CVG2 CV G1 CV
CVCV
CV
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1. RMSroot mean square errorRMS
2. G1G1
SS
G2G2
3. G1G1
G2G2
4. CVcoefficient of variationG0/G1
CVCV
G1 cvG1G1
G2 cvG2
Flowjo
FlowJo FlJ FlowJo
