Turner Quantech Digital Filter Fluorometer...6 Installation The Turner Quantech Digital Filter...

1LT1095X1 • 5/18/99

BARNSTEAD|THERMOLYNE CORPORATION

Turner® Quantech™Digital Filter Fluorometer

Operation Manual

Model # Description Voltage SeriesFM109515 Base Model, Wavelength Range 340 - 650 nm 120 Volts 1095FM109514 Base Model, Wavelength Range 340 - 650 nm 100 Volts 1095FM109510-33 Base Model, Wavelength Range 340 - 650 nm 230 Volts 1095

FM109525 IR Model, Wavelength Range 340 - 870 nm 120 Volts 1095FM109524 IR Model, Wavelength Range 340 - 870 nm 100 Volts 1095FM109520-33 IR Model, Wavelength Range 340 - 870 nm 230 Volts 1095

FM109535 Wide Band Model, Wavelength Range 254* - 870 nm 120 Volts 1095FM109534 Wide Band Model, Wavelength Range 254* - 870 nm 100 Volts 1095FM109530-33 Wide Band Model, Wavelength Range 254* - 870 nm 230 Volts 1095

* Wide band model contains a mercury line lamp that provides major emission lines at 254, 313, 360,405, 436, 546 and 578 nm.

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Table of ContentsIntroduction ........................................................................................................................................................................... 3Safety Information ................................................................................................................................................................. 4 Intended Use ....................................................................................................................................................................... 4 Warnings ............................................................................................................................................................................. 4Unpacking ............................................................................................................................................................................. 5Installation ............................................................................................................................................................................. 6Specifications ........................................................................................................................................................................ 7 Environmental Conditions ................................................................................................................................................. 12 Declaration of Conformity ................................................................................................................................................. 12General Description ............................................................................................................................................................ 13 Components ...................................................................................................................................................................... 13 Keypad and Function Keys ............................................................................................................................................... 14Operation ............................................................................................................................................................................ 18 Unit Power-Up ................................................................................................................................................................... 18 Performing Fluorescence Analysis .................................................................................................................................... 18 Aflatoxin, DNA, Rhodamine and Histamine Methods ........................................................................................................ 20 New Method ...................................................................................................................................................................... 24 Raw Fluorescence (Mode I) .............................................................................................................................................. 28 Raw Fluorescence (Mode II) ............................................................................................................................................. 29 Advanced Functions .......................................................................................................................................................... 31 Diagnostic Menu ............................................................................................................................................................... 32 Set Printer and Print Options ............................................................................................................................................ 32 Unit Power-Down .............................................................................................................................................................. 33 Standard Curve - Linear Regression ................................................................................................................................ 33 Gain Code Table ............................................................................................................................................................... 35 Verification of Linearity ...................................................................................................................................................... 36 Performance Verification ................................................................................................................................................... 36Printout Capability/Computer Connection ........................................................................................................................... 38 Connecting the Quantech Filter Fluorometer to Computer and Communicating Through the RS-232 Port UsingHyperterminal or Procomm ................................................................................................................................................. 38 Connecting and Starting the Printer .................................................................................................................................. 41Maintenance ....................................................................................................................................................................... 43 Excitation and Emission Filters ......................................................................................................................................... 43 Gelatin and Interference Coated Filters ............................................................................................................................ 43 Sample Chamber and Cuvette Holder Cleaning ............................................................................................................... 44Exploded View .................................................................................................................................................................... 45Replacement Parts ............................................................................................................................................................. 46 Gaining Access to Replacement Parts .............................................................................................................................. 46Appendix ............................................................................................................................................................................. 48 Theory of Fluorescence .................................................................................................................................................... 48 Fluorescence/Concentration Relationships ...................................................................................................................... 49 Considerations .................................................................................................................................................................. 50 Standards .......................................................................................................................................................................... 51 Glassware ......................................................................................................................................................................... 51 Temperature ...................................................................................................................................................................... 51 pH...................................................................................................................................................................................... 52 Miscellaneous ................................................................................................................................................................... 52 Cuvette Matching Procedure ............................................................................................................................................ 53 Excitation Sources ............................................................................................................................................................ 54 Filters ................................................................................................................................................................................ 55 Filter Selection Chart ........................................................................................................................................................ 60 Table I ................................................................................................................................................................................ 61 Table II ............................................................................................................................................................................... 62 Printer Setup ..................................................................................................................................................................... 65 Quantech Accessories ...................................................................................................................................................... 67 Flowchart ........................................................................................................................................................................... 69Ordering Procedures ........................................................................................................................................................... 77Two Year Limited Warranty ................................................................................................................................................. 80

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Introduction

The Turner Quantech Digital Filter Fluorometeris a highly sensitive, microprocessor-controlledinstrument utilizing integrated circuit analog anddigital signal processing. Excitation energy isprovided by a quartz-halogen lamp anddetection of the emission light is provided by ahigh gain, low noise photomultiplier tube. Theoperating wavelength range of the fluorometerdepends on the model purchased. Excitationand emission wavelength selection isaccomplished by filters.

Menu-driven software guides the user throughsetting up a multipoint (9 points maximum)standard curve for any given “Method.” Unit willstore standard curve data (2 points to 9 points)for up to 9 “Methods.” Gain selection isperformed automatically by the fluorometersoftware.

The Turner Quantech Digital Filter Fluorometeris designed to perform analytical quantitativefluorescence measurements on variousfluorescent materials including chlorophyll,fluorescein, histamine, vitamins, rhodamine,DNA/RNA dye complexes and other fluorescentcompounds.

The Turner Quantech Digital Filter Fluorometerwill automatically set optimal sensitivity andrange for your compounds/samples and displaya readout of the actual concentration of anunknown sample.

NotePlease read this instructionmanual carefully to ensureoptimal operation of the unit.

NoteThe Turner Quantech DigitalFilter Fluorometer is intended forresearch purposes only.

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Your Turner Quantech Digital Filter Fluorometerhas been designed with function, reliability andsafety in mind. It is the user’s responsibility toinstall it in conformance with local electricalcodes. For safe operation, please observe thealert signals throughout this manual.

Intended UseDo not use this product for anything other than itsintended use.

WarningsTo avoid electrical shock, always:

1. Use a properly grounded electrical outletof correct voltage and current handlingcapacity.

2. Disconnect from the power supply beforemaintenance and servicing.

To avoid personal injury:

1. Refer servicing to qualified personnel.

Safety Information

WarningWarnings alert you to apossibility of personal injury.

CautionCautions alert you to a possibilityof damage to the equipment.

NoteNotes alert you to pertinent factsand conditions.

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Unpacking

After the Turner Quantech Digital FilterFluorometer is unpacked, inspect it for physicaldamage. Report any observed damageimmediately to the Freight Carrier and yourdealer.

Standard Equipment:1 Turner Quantech Digital Filter

Fluorometer

1 Power Cord

1 Excitation Filter (NB360, part no.LE1095X30)

1 Emission Filter (SC415, part no.LE1095X18)

1 Blocking Filter (Included with WideBand models only; for use in blockingout one of two light sources.)

1 Operation Manual (part no. LT1095X1)

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Installation

The Turner Quantech Digital Filter Fluorometerrequires a properly wired and grounded electricaloutlet of correct voltage and current handlingcapacity.

1. Position the instrument on a dry,vibration-free laboratory surface. Allow forat least 2” of open space behind the backof the unit for air circulation and the powercord.

2. Locate the power entry module on theback panel of the unit and connect thepower cord. (Refer to Figure 1.)

3. Plug the power cord into the properelectrical outlet.

4. Refer to the Operation section of thismanual before powering up thefluorometer.

NoteThe power entry module alsohouses the fuse drawer/holder.The fuse drawer/holder islocated to the right of the mainpower switch when viewing theback panel of the unit. Twofuses are provided with the unit.

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Part Number: FM109524 (IR Model)

Configuration: Filter Fluorometer (Filters sold separately)

Display: 2 x 24 LCD

Wavelength Range: Excitation: 340 - 750 nm

Light Source: Quartz Halogen Lamp 6V, 20W (Lamp life: 10,000 hours)

Emission: 185 - 870 nm

Detector: 931B PMT

Wavelength Accuracy: 1.5 - 10 nm (Depends on filters selected)

Bandwidth: 5 - 200 nm (Depends on filters selected)

Linearity: 1.00%

Sensitivity: 25 pg/mL dsDNA w/ PicoGreen Reagent, 30 ppt QSO4, 20 ppt FITC, 30 pg/uL ATP Luminescence

Concentration Range: 0.00 - 9999.99 (ppt, ppm, ppb, ug/dL, ug/mL, ng/mL, pg/mL, ng/uL)

Gain & Range Select: Automatic, 6 Orders

Menu Driven Software: Std. Curve Storage, 2 to 9 point Std. Curves, Store up to 9 Curves, Blank / Zero Set, Raw Fluorescence,

Discrete Signal Averaging, Diagnostics Menus, Injector Option

Languages (Selectable): English/Spanish/French/German

Data Output: RS-232 Serial Interface for Printer or Computer (9 pin), Optional Analog (0 - 1 VDC)

Sample Holders: 12.5x12.5x45 mm sq.cuvettes (adj. z dim. 8.5 or 15 mm), 12 x 75 mm round, Capillary Tube Option,

Flow Cell Option

Operating Temp. Range: 20° to 38°CPower: 100 V, 50/60 Hz

Dimensions: 27 cm W X 38 cm D X 16 cm H

Product Approval: CSA/NRTL/C

Optional Accessories: Epson™ Printer

Weight: 5.9 kg

QSO4 = Quinine Sulfate & FITC = Fluorescein Isothiocyanate • PicoGreen is a trademark of Molecular Probes, Inc.

Part Number: FM109514 (Base Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


Specifications

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SPECIFICATIONS

Part Number: FM109515 (Base Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


Part Number: FM109534 (Wide Band Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


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SPECIFICATIONS

Part Number: FM109535 (Wide Band Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


Part Number: FM109525 (IR Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


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SPECIFICATIONS

Part Number: FM109520-33 (IR Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option



Product Approval: CSA/NRTL/C, CE


Weight: 5.9 kg


Part Number: FM109510-33 (Base Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


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SPECIFICATIONS

Part Number: FM109530-33 (Wide Band Model)


Display: 2 x 24 LCD




Detector: 931B PMT



Linearity: 1.00%









Flow Cell Option





Weight: 5.9 kg


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Environmental ConditionsOperating: 20°C to 38°C; 20% to 80% relative

humidity, non-condensing. Installationcategory II (overvoltage) in

accordance with IEC 664. Pollutiondegree in accordance with IEC 664.Altitude Limit: 2,000 meters.

Storage: -25°C to 65°C10% to 85% relative humidity

Declaration of ConformityBarnstead/Thermolyne hereby declares under its soleresponsibility that this product conforms with the technicalrequirements of the following standards (230 volt/ -33models only):

EMC: EN 61000-3-2 Limits for HarmonicCurrent Emissions

EN 61000-3-3 Limits for Voltage Fluctuationsand Flicker

EN 61326 Electrical Equipment forMeasurement, Control andLaboratory Use; Part I: GeneralRequirements

Safety: EN 61010-1 Safety Requirements forElectrical Equipment forMeasurement, Control andLaboratory Use, Part I:General Requirements

per the provisions of the Low Voltage Directive 73/23/EEC, as amended by 93/68/EEC.

The authorized representative located within theEuropean Community is:

Electrothermal Engineering Ltd.419 Sutton RoadSouthend On SeaEssex SS2 5PHUnited Kingdom

Copies of the Declaration of Conformity are availableupon request.

SPECIFICATIONS

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ComponentsDisplay: 2 x 24 character LCD display toindicate software menus and fluorescencevalues.

Power Switch: rocker switch located on backpanel. (Refer to Figure 1.)

Sample Chamber and Cuvette Holder: thelarge sample chamber will accommodatenonstandard size cuvettes and up to four flowtubes. The standard cuvette holder suppliedwith the units accepts 12 x 75 mm roundcuvettes and 12.5 x 12.5 x 45 mm squarecuvettes. To insert the cuvette in the holder,open the chamber cover and carefully insert thecuvette by positioning it between the spring clipand the angular sides of the holder. Two flat,clear faces of a square cuvette should opposethe circular openings that form the light paths.Take care not to touch the curved mirror whichis located next to the spring clip (refer to Figure2).

To Accommodate Small Sample Volumes(2ml or less) in Standard Cuvettes: Removethe two Phillips head screws holding thecuvette holder in place, carefully lift the holderout of the sample chamber (avoid touching thecurved mirror) and press up on the round, blackcylinder located on the bottom of the holder.Carefully reinsert the cuvette holder into thesample chamber using the two mountingscrews.

Filter Block and Filter Slides: The filter blocklines the perimeter of the sample and serves asa holder for the Excitation and Emission filterslides. The Visible Range Excitation filter slide

NoteThe Spring clip in the cuvetteholder is adjustable and can bepushed inward to accommodate 10mm diameter tubes or outward toaccommodate 16 mm diametertubes. (Maximum tube height is130 mm 13 cm).

NoteThe z dimension of the samplechamber is 15 mm when the blackcylinder is in the “normal” downposition. When the black cylinder isin the up position, the samplechamber z dimension is 8.5 mm.

CautionDo not touch the curved mirrorlocated in the cuvette holder.

General Description

Figure 1: Back View

Serial Data Port(connect to printer or computer)

Power EntryModule

Port for Optional InjectorPump Control Cable

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is placed in the trapezoidal-shaped holder locatedon the back side of the sample chamber (refer toFigure 2). The Emission filter slide is placed in thetrapezoidal-shaped holder located on the left sideof the sample chamber (refer to Figure 2).

To Change the Filter Slide: Open the chambercover and grip the top of the filter slide with yourthumb and index finger so you can lift the slide upand out of the filter slide holder. When replacing afilter slide be sure that the circular filter is properlyaligned with the light path opening in the bottomof the filter holder.

Keypad and Function Keys(See Figure 3.)

UP ARROW (ON/YES) key: Allows operator toscroll through “Method” parameters in anincremental manner and select an option or selectthe “ON/YES” option in response to a questionposed by the fluorometer software user interface.

DOWN ARROW (OFF/NO) key: Allows operatorto scroll through “Method” parameters indescending order and select an option or selectthe “OFF/NO” option in response to a questionposed by the fluorometer software user interface.

LEFT ARROW key: Allows operator to scrollbetween “Menu,” “Method” and “Function” optionsand select an option, or scroll across the display.

RIGHT ARROW key: Allows operator to scrollbetween “Menu,” “Method” and “Function” optionsand select an option, or scroll across the display.

CautionUse finger pressure only whendepressing keys on keypad;depressing keys with pens orother utensils may causepermanent damage to thekeypad.

GENERAL DESCRIPTION

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ENTER key: Allows operator to activate orproceed with a selected option visible on thedisplay screen.

MENU key: Serves as a home key to allowoperator to immediately return to the “MainMenu.”

BACK key: Allows operator to go back one stepwithin a “Method,” “Menu” or “Function.”

PRINT key: Initiates the signal to send standardcurve data, sample concentration data andmethod settings to a printer or computer.

ZERO KEY: Sets fluorescence value to zerowhen blank solution is in the cuvette holder.

GENERAL DESCRIPTION

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GENERAL DESCRIPTION

Chamber Cover

Insert Visible Range Excitation Filter HereAn Excitation filter placed in this positionfilters the quartz-halogen light source in allQuantech fluorometer models.

Insert Emission Filter HereThe Emission filter passes desiredfluorescence wavelengths to be detectedby the photomultiplier tube (PMT).

Sample Chamber/Cuvette Holder

Insert UV Range Excitation Filter HereAn Excitation filter placed in this holder filters themercury line lamp (Light source) in the Wide Bandmodel of the Quantech fluorometer.

Spring Clip

Figure 2

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GENERAL DESCRIPTION

Display

Keypad

ENTER key

UP ARROW(ON/YES) key

DOWN ARROW(OFF/NO) key

MENU key BACK key PRINT key

LEFT ARROW key

RIGHT ARROW key

ZERO key

Figure 3

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Unit Power-Up1. Turn main power switch located on

back of unit ON.

2. Display reads, “Turner Quantech Fluo-rometer Manufactured by Barnstead/Thermolyne Fluorometer, SoftwareVersion 1.00.03.”

3. Display proceeds to the Languageselection option. The default languageis “English.” To select another language(Espanol, French, Deutsch), press theLEFT or RIGHT ARROW key.

4. Press ENTER to continue operation ofthe fluorometer.

5. Display reads, “Unit Initializing, PleaseWait;” “Auto-Calibrating (Please Wait);”“Unit Initializing, “15 Minutes Remain.”

6. The unit will perform a 15 minute warm-up countdown to ensure the stability ofthe fluorometer electronics. The count-down timer on the display allows you tomonitor the time remaining before thewarm-up period is complete.

Performing FluorescenceAnalysisAfter the unit completes its warm-up, thedisplay will rest at the “Main Menu”. At thispoint the user can perform “AdvancedFunctions” or choose a method of analysis.

Operation

NoteThe English language option is thedefault language. Operator mayselect Espanol, French or Deutsch.

NoteBefore performing fluorescenceanalysis, please consult theAppendix of this manual forinformation on cuvettes, filterselection and the theory offluorescence.

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The operation of the Base, IR and Wide Bandmodels of the Quantech fluorometers are identicalwith the exception of the option to activate the UVlamp (mercury line lamp) for applicationsrequiring excitation wavelengths below 340 nm.

Advanced FunctionsPress the LEFT or RIGHT ARROW key to selectthe desired option:

• Date and Time

• View Stored Data

• Reset System to Default

• View Diagnostic Information

• Set Printer and Print Options

• UV Lamp Options

• Manually Set Gain and PMT Voltages

• Injector Pump Options.

Choosing a Method of Analysis1. Starting from the “Main Menu,” press the

ENTER key until display reads, “ChooseMethod.”

2. Press the LEFT or RIGHT ARROW key tochoose from the following methods:

• Aflatoxin• DNA• Histamine• Rhodamine• New Program• Raw Fluorescence

3. Select the desired method and pressENTER.

OPERATION

NotePlease refer to the AdvancedFunctions section of thismanual and the flowchartlocated in the Appendix fordirections on how to operateand utilize these options.

NoteManual settings will beoverridden if you choose astored curve.

NotePressing the MENU key willreturn you to the “Main Menu”home setting. Pressing theBACK key will allow you to goback one step.

NoteThe Aflatoxin, DNA, Histamineand Rhodamine methods serveas memory location holders.New Quantech fluorometers donot include standard curves forthese methods. These methodswere chosen as memorylocation holders because theyare common fluorescencemethods.

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Follow the proceeding steps according to themethod selected.

Aflatoxin, DNA, Rhodamine andHistamine MethodsRefer to the flowchart in the Appendix for furtherinformation.

After choosing either the Aflatoxin, DNA,Rhodamine or Histamine method, proceed withthe following steps:

1. Display reads, “Change Name? (Y/N?).”

a. Press ENTER to select “No.” (Systemwill automatically default to “No.”)Proceed to step 2.

b. If you wish to change the “Method”name, press the UP ARROW key toselect the “Yes” option and pressENTER. See the New Method sectionfor instructions on how to change themethod name.

2. Display reads, “Proper Filters in Unit?”“Yes.” (System defaults to “Yes.”)

a. If you are satisfied with the filters thatyou have in the unit for your method ofchoice, press the ENTER to select“Yes” and proceed to step 3.

b. If you need help choosing the correctExcitation and Emission filter slides foryour method, press the DOWN AR-ROW key to select the “No” option andpress ENTER. Display will first indicatea method option. To scroll between themethods for which selection information

OPERATION

NoteChanging a program name willautomatically delete the datafrom the previous method.

NoteIf your method of fluorescenceanalysis requires the mercuryline lamp as a light source (WideBand model only) activate theUV lamp in the “AdvancedFunctions” (UV Lamp Options)before proceeding with yourmethod of analysis. Rememberto place the blocking filterincluded with the QuantechWide Band model in the VisibleRange Excitation Filter holder(see Figure 1).

To use your Wide Band modelover 340 nm, place the blockingfilter in the UV Range Excitationfilter holder and deactivate theUV lamp in “AdvancedFunctions.”

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is available, press the LEFT andRIGHT ARROW keys. When thedisplay indicates the desired methodoption, press the UP ARROW key toobtain a recommendation for the“Primary Excitation Filter” and thenpress the DOWN ARROW key toobtain a recommendation for the“Secondary Emission Filter.” Toreturn to setting up your method,press ENTER.

3. Display reads, “Std Curve from Memory(Y/N?); No.” (System will automaticallydefault to “No.”)

a. If no, press ENTER and proceed tostep 4.

b. If yes, press the UP ARROW toselect “Yes” and press ENTER.Proceed to step 8.

4. Display reads, “Enter Number ofPoints;” “2.” The default setting for thenumber of points in a standard curve is2. Enter a value for the number ofstandards of known concentration thatyou will be working with by pressing theUP and DOWN ARROW keys to scrollbetween numbers in positive and nega-tive increments, respectively. PressENTER.

5. Display reads, “Enter Concentration;”“0000.000 <FIU>.” Press LEFT orRIGHT ARROW key to scroll horizon-tally across the display and select acharacter. Each “0” represents anumerical character field that can be

OPERATION

NoteThe Turner Quantech DigitalFilter Fluorometer will store yourstandard curve data in memorypermanently. Memory capacitywill allow a total of 9 methodswith 9 points.

NoteNew fluorometers do not containstored standard curve data forany methods.

NoteRemember to input the correctvalue and units for the standardwith the maximum concentrationfirst. For any given standardcurve, only one unit of measurecan be used and is set when theunit of measure for the firststandard is entered.

NoteThe blank solution should not beincluded as a point in the stan-dard curve.

Note<FIU> indicates a generic unit ofmeasure or fluorescence Inten-sity units.

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changed. The <FIU> field can also bechanged when the “>” is highlighted. Inorder to select a concentration valueand unit of measure, press the UP andDOWN ARROW keys to scroll betweennumbers and concentration units (ppm,ppb, ppt, µg/ml, µg/dl, ng/ml, pg/ml orng/µl). When the display reflects thecorrect numerical value and units forthe first standard (known sample),Press ENTER.

6. Display reads, “Insert Known Sample#1.” Open the chamber cover, insertsample and close cover. Press ENTER.Display then reads, “Auto Gain Setting;”Please Wait” and runs through con-secutive gain settings before showing,“Taking Reading.”

Next the display reads, “Enter Concen-tration;” “0000.00 <FIU>.” Select a newconcentration value and press ENTER.Display reads, “Insert Known Sample2.” Open the chamber cover, insertknown sample #2 and close the cover.Press ENTER. Display will read, “Tak-ing Reading.” Repeat these steps foreach known sample in the standardcurve.

7. Display reads, “Insert Blank.” Openchamber cover and insert cuvette withblank solution in the cuvette holder.Close chamber cover and press ZERO.The display will read, “Taking Reading”and then show the “Coefficient of De-termination;” “1.00” (example value).Proceed to step 8.

OPERATION

NoteAfter known sample #1, the maxi-mum concentration is measured,the remaining known samples canbe inserted in any order to com-plete the standard curve for yourmethod. It is recommended toinsert known samples in decreas-ing order of concentration.

NoteIMPORTANT: Pressing BACK afterthe unit has taken a reading for aknown sample in the standardcurve will take you back to “EnterNumber of Points; “2” and you willneed to repeat the process ofmeasuring known samples from #1on. The BACK key can be usedwhen a correction to a concentra-tion value is needed provided thatENTER has not been pressed.

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If an unacceptable coefficient (<0.75) isobtained, the unit will display the valueand indicate “Curve Unacceptable;” PressEnter.” After you press ENTER, the dis-play will indicate “Enter Number ofPoints;” “2.” Please check the concentra-tions of your standard samples beforerepeating the steps of the standard curveprocedure.

8. If an acceptable “Coefficient of Determi-nation” is obtained, the display will read“Insert Unknown Sample.” After insertingthe cuvette containing the unknownsample into the cuvette holder and clos-ing the chamber cover press ENTER. Thedisplay will read, “Taking Reading” andthe concentration value of the unknownsample is displayed. The standard curvedata for your method is now stored inmemory. A concentration reading of thefirst unknown sample is required to storethe standard curve data.

To continue fluorescence analysis of unknownsamples, remove the previous sample for whichthe concentration value was determined andinsert the next unknown sample. After closing thechamber cover, press ENTER to obtain theconcentration value of the sample. Repeat thesesteps for all unknown samples.

If the concentration of an unknown sample doesnot fall within the range of your standard curve,the unit will display an “errant value” or indicate“range error.” A new standard curve will need tobe set up in order to correctly analyze an over orunder range sample. Alternatively, the unknownsample can be diluted by a given factor and thedilute sample analyzed.

OPERATION

NoteThe standard curve data thatyou have just obtained can beutilized for future analyses.However, please note that it isgood laboratory practice toperform new standard curves atleast once a week. TheQuantech fluorometer will keeptrack of the age of your standardcurve data. After 5 days the unitwill indicate that the curveshould be considered marginalby prompting you on whether ornot (YES or NO) to use the databefore proceeding with analysis.

NoteIf you are using a standardcurve from the memory of theQuantech unit, the display willfirst indicate the age of thestandard curve data and thecoefficient of determination forthe curve. The display will thenread, “Insert Unknown Sample.”

NoteThe blank for a stored standardcurve may be reset to zero.When resetting the blank to zerousing the ZERO key, you arereplacing the last blank settingwith the new blank setting. Thisreset procedure will not com-pensate for errors in samplereadings due to instrument drift.For maximum accuracy it is bestto redo your curve.

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New Method1. Display reads, “New Method (Y/N?);

No.” (System will automatically defaultto “No.”) and provides an alphabetical/character listing. Alternatively, thedisplay will read “New Program” andprovide the same options as “NewMethod.”

2. To change the method name, press theLEFT and RIGHT ARROW keys toselect a letter or character from thealphabetical/character listing on thedisplay. Each time an ARROW key ismanually depressed, the display cursorwill advance or step back one charac-ter. After selecting a character (cursorhighlights the character), press the UPARROW key. The letter you haveselected will appear at the top of thedisplay window. Continue selectingletters in this manner until you haveentered the desired program name,then press ENTER.

3. Display reads, “Proper Filters in Unit?”;“Yes” (System defaults to “Yes.”)

a. If you are satisfied with the filtersthat you have in the unit for yourmethod of choice, press ENTER toselect “Yes” and proceed to step 3.

b. If you need help choosing the correctExcitation and Emission filter slidesfor your method, press the DOWNARROW key to select “No” and press

OPERATION

NoteIf you select a character in error,you can backspace or delete thecharacter by pressing theDOWN ARROW key. Afterdeleting a character, select anew character by pressing theLEFT or RIGHT ARROW key.

NoteNew Method offers the option tocustomize the methods that arestored in the Quantech Fluorom-eter.

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ENTER. Display will first indicate amethod option. To scroll between themethods for which selection informationis available, press the LEFT andRIGHT ARROW keys. When the dis-play indicates the desired methodoption, press the UP ARROW key toobtain a recommendation for the “Pri-mary Excitation Filter” and then pressthe DOWN ARROW key to obtain arecommendation for the “SecondaryEmission Filter.” To return to setting upyour method, press ENTER.

4. Display reads, “Std Curve from Memory(Y/N?); No.” (System will automaticallydefault to “No.”)

a. If no, press ENTER and proceed tostep 5.

b. If yes, press the UP ARROW to select“Yes” and press ENTER. Proceed tostep 9.

5. Display reads, “Enter Number of Points;”“2.” The default setting for the number ofpoints in a standard curve is 2. Enter avalue for the number of standards ofknown concentration that you will beworking with by pressing the UP andDOWN ARROW keys to scroll betweennumbers in positive and negative incre-ments, respectively. Press ENTER.

6. Display reads, “Enter Concentration;”“0000.000 <FIU>.” Press the LEFT orRIGHT ARROW key to scroll horizontallyacross the display and select a characterthat is to be changed. Each “0” represents

OPERATION

NoteThe Turner Quantech DigitalFilter Fluorometer will store yourstandard curve data in memorypermanently. Memory capacitywill allow a total of 9 methodswith 9 points.

NoteNew fluorometers do not containstored standard curve data forany methods.

NoteRemember to input the correctvalue and units for the standardwith the maximum concentrationfirst. For any given standardcurve, only one unit of measurecan be used and is set when theunit of measure for the firststandard is entered. <FIU>indicates a generic unit ofmeasure or FluorescenceIntensity Units.

NoteThe blank solution should not beincluded as a point in the stan-dard curve.

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a numerical character field that can bechanged. The <FIU> field can also bechanged when the “>” is highlighted. Inorder to select a concentration valueand unit of measure, press the UP andDOWN ARROW key to scroll betweennumbers and concentration units (ppm,ppb, ppt, µg/dl, µg/ml, ng/ml, pg/ml orng/µl). When the display reflects thecorrect numerical value and units forthe first standard (known sample),Press ENTER.

7. Display reads, “Insert Known Sample#1.” Open the chamber cover, insertsample and close cover. Press ENTER.Display then reads, “Auto Gain Setting;”“Please Wait” and runs through con-secutive gain settings before showing“Taking Reading.”

Next the display reads, “Enter Concen-tration;” “0000.00 <FIU>.” Select a newconcentration value and press ENTER.Display reads, “Insert Known Sample2.” Open the chamber cover, insertknown sample #2, and close the cover.Press ENTER. Display will read, “Tak-ing Reading.” Repeat these steps foreach known sample in the standardcurve.

8. Display reads, “Insert Blank.” Openchamber cover and insert cuvette withblank solution in the cuvette holder.Close chamber cover and press ZERO.The display will indicate “Taking Read-ing” and then show the “Coefficient ofDetermination;” “1.00” (example value).Proceed to step 9.

OPERATION

NoteRemember to input the correctvalue and units for the standardwith the maximum concentrationfirst. For any given standardcurve, only one unit of measurecan be used and is set when theunit of measure for the firststandard is entered.

NoteAfter known sample #1, themaximum concentration ismeasured, the remaining knownsamples can be inserted in anyorder to complete the standardcurve for your method. It isrecommended to insert knownsamples in decreasing order ofconcentration.

NoteIMPORTANT: Pressing BACKafter the unit has taken a read-ing for a known sample in thestandard curve will take youback to “Enter Number ofPoints; “2” and you will need torepeat the process of measuringknown samples from #1 on. TheBACK key can be used when acorrection to a concentrationvalue is needed provided thatENTER has not been pressed.

27

If an unacceptable coefficient (<0.75) isobtained, the unit will display the valueand indicate “Curve Unacceptable;”“Press Enter.” After you press ENTER,the display will indicate “Enter Number ofPoints;” “2.” Please check the concentra-tions of your standard samples beforerepeating the steps of the standard curveprocedure.

9. If an acceptable “Coefficient of Determi-nation” is obtained, the display will read“Insert Unknown Sample.” After insertingthe cuvette containing the unknownsample into the cuvette holder and clos-ing the cover press ENTER. The displaywill read, “Taking Reading” and the con-centration value of the unknown samplewill be displayed. The standard curvedata for your method is now stored inmemory. A concentration reading of thefirst unknown sample is required to storethe standard curve data.

To continue fluorescence analysis ofunknown samples, remove the previoussample for which the concentration valuewas determined and insert the next un-known sample. After closing the chambercover, press ENTER to obtain the con-centration value of the sample. Repeatthese steps for all unknown samples.

If the concentration of an unknownsample does not fall within the range ofyour standard curve, the unit will displayan “errant value” or indicate “range error.”A new standard curve will need to be setup in order to correctly analyze an over orunder range sample. Alternatively, theunknown sample can be diluted by agiven factor and the diluted sample ana-lyzed.

OPERATION

NoteThe standard curve data thatyou have just obtained can beutilized for future analyses.However, please note that it isgood laboratory practice toperform new standard curves atleast once a week.

NoteIf you are using a standardcurve from the memory of theQuantech unit, the display willindicate the age of the standardcurve data and the coefficient ofdetermination for the curve. Thedisplay will then read, “InsertUnknown Sample.”

NoteThe blank for a stored standardcurve may be reset to zero.When resetting the blank to zerousing the ZERO key, you arereplacing the last blank settingwith the new blank setting. Thisreset procedure will not com-pensate for errors in samplereadings due to instrument drift.For maximum accuracy it is bestto redo your curve.

28

Raw Fluorescence (Mode I)1. Display reads, “Enter Concentration;”

“000.000 <FIU>.” Press the LEFT orRight ARROW key to scroll horizontallyacross the display and select a charac-ter field that is to be changed. Each “0”represents a numerical character fieldthat can be changed. The <FIU> fieldcan also be changed when the “>” ishighlighted. In order to select a concen-tration value and unit of measure foryour reference sample, press the UP orDOWN ARROW key to scroll betweennumbers and concentration units (ppm,ppb, ppt, µg/ml, µg/dl, ng/ml, pg/ml, orng/µl). When the display reflects thecorrect value and units for the refer-ence sample, press ENTER.

2. Display reads, “Insert Known Sample;”Open the chamber cover, insert refer-ence sample and close the cover.Press ENTER. Display then reads,“Auto Gain Setting;” Please Wait” andruns through consecutive gain settingsbefore showing “Taking Reading.”

3. Display reads, “Insert Blank;” “No”System defaults to “No.”

If you choose to use a blank sample in“Raw Fluorescence,” press the UPARROW key to select “Yes.” Displayreads, “Insert Blank Sample.” Open thechamber cover, insert blank sampleand close the cover. Press ZERO.Display then reads, “Taking Reading.”Proceed to step 4.

OPERATION

Note<FIU> indicates a generic unit ofmeasure or Fluorescence Inten-sity Units.

NoteRaw Fluorescence (Mode I) isdesigned for applications that donot require 2 - 9 point standardcurves. This mode essentiallyprovides a 1 point standardcurve in reference to zero con-centration of a compound. This1 point is only stored in memoryfor the duration of your rawfluorescence analysis.

NoteA reference sample is generallyof the highest concentration youwould expect to analyze for thecompound of interest. An arbi-trary numerical value is usuallyassigned as the concentration ofthis reference sample (e.g.1000.000 <FIU> or 1000.000ppb.

29

If you do not want to use a blank sample,press the ENTER key to select “No” andproceed to step 4.

4. Display reads “Insert Unknown Sample.”Open the chamber cover, insert unknownsample, close cover and press ENTER.The display will show the “Raw Concen-tration” value of the sample per the refer-ence sample unit of measure.

5 To continue determination of “Raw Con-centration” values for subsequent un-known samples, open the chamber cover,insert next unknown sample and closecover. The display will show the “RawConcentration” of the unknown samplewithin approximately two seconds.

6. If the unknown sample is significantlymore fluorescent than the referencesample, the display will indicate an over-range error. The unknown sample can bediluted by a given factor and read againor the unknown can serve as the newreference sample in a raw fluorescenceanalysis.

Raw Fluorescence (Mode II)1. Display reads, “Enter Concentration;”

“0000.000 <FIU>.” Press ENTER.

2. Display reads, “Insert Known Sample.”Open the chamber cover, insert referencesample and close the cover. Press EN-TER. Display then reads, “Auto GainSetting;” Please Wait” and runs throughconsecutive gain settings before showing“Taking Reading.”

OPERATION

Note<FIU> indicates a generic unit ofmeasure or FluorescenceIntensity Units.

30

3. Display reads, “Insert Blank;” “No.” (Sys-tem defaults to “No.”)

If you choose to use a blank sample in“Raw Fluorescence,” press the UP AR-ROW key to select “Yes.” Display reads,“Insert Blank Sample.” Open the cham-ber cover, insert reference sample andclose the cover. Press ZERO. Displaythen reads, “Taking Reading.” Proceed tostep 4.

If you do not want to use a blank sample,press the ENTER key to select “No” andproceed to step 4.

4. Display reads “Insert Unknown Sample.”Open the chamber cover, insert unknownsample, close cover and press ENTER.The display will indicate the “Raw Con-centration” value of the sample per thereference sample unit of measure.

5 To continue determination of “Raw Con-centration” values for subsequent un-known samples, open the chamber cover,insert next unknown sample and closecover. The display will show the “RawConcentration” of the unknown samplewithin approximately two seconds.

6. If the unknown sample is significantlymore fluorescent than the referencesample, the display will indicate an over-range error. The unknown sample can bediluted by a given factor and read againor the unknown can serve as the newreference sample in a raw fluorescenceanalysis.

OPERATION

NoteRaw Fluorescence (Mode II) isdesigned for basic researchapplications in which a refer-ence standard of “known” con-centration is not available. TheTurner Quantech fluorometerassigns a value to a referencesample based upon the autogain function response. Ingeneral, the reference samplewould be chosen such that itrepresents the maximum fluo-rescence response expected inthe range of unknown samplesthat are to be analyzed. The“Raw Fluorescence” of theunknown samples is determinedrelative to the reference sample.The fluorescence response forthe reference sample is onlystored in memory for the dura-tion of your “Raw Fluorescence”analysis.

31

Advanced FunctionsTo access the “Advanced Functions” parameterfrom the “Main Menu,” press the LEFT or RIGHTARROW key until the display reads, “AdvancedFunctions,” then press ENTER. Press the LEFT orRIGHT ARROW key to step through the followingoptions:

• Set Date and Time: User can define month,day, year, hour, minutes and seconds.

• View Stored Data: User can view standardcurve data for defined methods.

• Reset System to Default: User can resetthe system to default settings in the rareevent that the RAM is corrupted.

• View Diagnostic Information: User canperform a status check on fluorometercomponent functions to aid in troubleshoot-ing.

• Set Printer and Print Options: User canset data output configurations for optionalprinter or computer communication.

• UV Lamp Options: User can activate theUV lamp (mercury line lamp) on the WideBand model of the Quantech.

• Manually Set Gain and PMT Voltages:User can turn off the auto gain function andmanually select gain and PMT Voltage.

• Injector Pump Options: User can activateoptional injector pump for kinetics studies.

Please refer to the flowchart in the Appendix foradditional navigational information.

NoteResetting the system will resultin loss of all standard curves.

OPERATION

32

Diagnostic MenuThe diagnostic menu within “AdvancedFunctions” is designed to help the operator ofthe Quantech fluorometer perform statuschecks on the components and systems of theunit. In the event that the unit malfunctions, thevalues indicated for the PMT, lamps, and powersupply will provide you and/or a Barnstead/Thermolyne technical representative with theinformation necessary to troubleshoot therepair of the unit.

To access the diagnostic menu from the “MainMenu,” press the LEFT or RIGHT ARROW keyuntil display reads, “Advanced Functions.”Press ENTER. Press the LEFT or RIGHTARROW key until the display reads, “ViewDiagnostic Information.” Press ENTER. Pressthe LEFT ARROW key to check on the statusof the PMT (e.g. raw PMT counts and A to Dcounts for the PMT at each Gain setting). Pressthe RIGHT ARROW key to view the “Hardware”e.g. Halogen lamp ON/OFF status and coverOPEN/CLOSED status). Press the ARROWkeys to step through the menu options. Pleaserefer to the flowchart in the Appendix foradditional navigational information for thesemenus.

Set Printer and Print OptionsExternal devices such as a serial printer or acomputer can be connected to the Quantechfluorometer via the RS-232 port on the back ofthe unit. Please refer to the printer section ofthe Appendix for instructions on obtaining aprintout with a printer or computer.

OPERATION

NoteThe left and right hand arrowsvisible in the display indicate thatnavigation options are availableupon pressing the LEFT or RIGHTARROW key.

33

OPERATION

Unit Power-DownUpon completing all fluorescence analyses withthe Turner Quantech Digital Filter Fluorometer,power down the unit by pressing the MENU keyto return to the “Main Menu.” Turn the main powerswitch located on the back panel of the unit OFF.

Standard Curve – LinearRegression

Calculation of Sample Concentrations

When you set up a standard curve using 2 to 9points, the Quantech software utilizes a linearregression routine that fits the concentration vs.fluorescence intensity response to the equation:y = mx + b.

Where: y = fluorescence intensityx = concentrationm = slope of the straight lineb = y axis intercept

Assuming that all standards utilized in a given“Method” are within the linear range of thefluorometer, a plot of standard concentration vs.fluorescence intensity will result in a straight line.

The Quantech Fluorometer linear regression doesnot utilize the blank sample as a point. Instead,because the unit allows the operator to reset thezero point for a given set of data associated witha specific method if they so choose, thefluorescence intensity of the blank sample issubtracted from the fluorescence intensity valuesthat define the standard curve in a separateoperation. The Quantech fluorometer calculates

34

the concentration of an unknown aftersubtracting the fluorescence contributed by theblank sample from the fluorescence associatedwith the unknown sample.

Viewing DataAn operator can examine the data used todefine the standard curve constructed for agiven “Method.” In the “Advanced Functions”menu (see flowcharts located in theAppendix), you can enter the “View StoredData” option. The Quantech fluorometer storesthe gain code determined by the auto gainfunction of the unit, the individualconcentrations of all standards used toconstruct a standard curve, the A to D readingsfor each standard which represent thephotomultiplier tube response to the standardconcentrations, the regression slope,regression intercept, and R-squared value orcoefficient of determination.

The data provided for each stored standardcurve allows the operator to determine thequality of the data immediately without havingto manually construct a concentration vs.fluorescence plot. This data is also stored inmemory for subsequent use until it is replaced.

The gain code indicated in the “View StoredData” option is designed to help the operatorgauge variations in standard and sampleconcentrations. The Gain Code of theQuantech fluorometer indicates the relativesensitivity of the response of the unit to thecompound under test (refer to Gain CodeTable).

OPERATION

35

Gain Code Table

Turner Quantech Fluorometer Autogain Function

GAIN CODE 1 = HIGHEST SENSITIVITYGAIN CODE 17 = LOWEST SENSITIVITY

GAIN CODE LAMP LEVEL SENSITIVITY PMT LEVEL1 HALOGEN 1 GAIN 1000 HV4 = HI2 HALOGEN 1 GAIN 1000 HV3 = MH3 HALOGEN 1 GAIN 100 HV4 = HI4 HALOGEN 1 GAIN 100 HV3 = MH5 HALOGEN 1 GAIN 10 HV4 = HI6 HALOGEN 1 GAIN 1000 HV2 = ML7 HALOGEN 1 GAIN 10 HV3 = MH8 HALOGEN 1 GAIN 1 HV4 = HI9 HALOGEN 1 GAIN 100 HV2 = ML10 HALOGEN 1 GAIN 1 HV3 = MH11 HALOGEN 1 GAIN 10 HV2 = ML12 HALOGEN 1 GAIN 1000 HV1 = LO13 HALOGEN 1 GAIN 1 HV2 = ML14 HALOGEN 1 GAIN 100 HV1 = LO15 HALOGEN 1 GAIN 10 HV1 = LO16 HALOGEN 1 GAIN 1 HV1 = LO17 HALOGEN 0 GAIN 1 HV1 = LO

Key for PMT Level Codes

HI HIGHMH MEDIUM HIGHML MEDIUM LOWLO LOW

OPERATION

36

Verification of LinearityThe linear range is the concentration range inwhich the LCD reading of the Quantech unit isdirectly proportional to the compoundconcentration. The linear range begins with thesmallest measurable concentration and spansto a maximum concentration limit that isdependent upon the chemical properties of thefluorescent compound under study, the filtersused, and the path length of the samplecuvette. Above the maximum concentrationlimit, the fluorescence readings level off or donot increase to the extent expected. Underthese circumstances the linear response limit ofthe compound has been reached. A newstandard curve is required for accuratereadings. At even higher concentrations abovethe maximum concentration limit, thefluorescent response of the fluorometer willactually begin to decrease even though thesample concentration is increasing. Thisphenomenon is known as “quenching.”

Performance VerificationIn the event that you are experiencing nonlinearresults with the compound that you areanalyzing and you want to verify theperformance of the Quantech unit, we suggestyou try a standard curve determination forQuinine Sulfate.

Quinine Sulfate Standard Reference Material(SRM) 936a may be obtained from the NationalInstitute for Standards and Technology (NIST),Standard Reference Material Program, Room204, Building 202, Gaithersburg, MD, 20899.Phone: 301-975-6776

OPERATION

37

OPERATION

Prepare a set of standard solutions of QuinineSulfate in 0.1 N Sulfuric Acid. A concentrationrange of 1 ppb to 100 ppb can be used. Usingthe NB360 filter for excitation and the SC415 filterfor emission, set up a 5 point standard curve withthe Quantech unit. A typical response is shown inFigure 4. Please note that it is critical to observegood laboratory practice when preparing standardsolutions. All glassware should be cleaned anddried.

Concentration (ppb) A/D Reading A/D Blank100 594 56980 476 45150 307 28220 137 1121 30 50 25 0

Linear Regressionslope 5.684574976intercept 23.4343362R-squared 0.999951429

Figure 4

38

Connecting Quantech FilterFluorometer to Computer andCommunicating Though theRS-232 Port UsingHyperterminal or Procomm

Hyperterminal

RS-232 Capture Instructions1. Connect the 9 pin serial cable (part no.

WHX18) from the Quantech fluorom-eter RS-232 port to COM2: port or(COM1: port) on back of computer

2. Access “Set Printer and Print Options”in the “Advanced Functions” menu (seeflowchart in Appendix). You may setyour print interval to a predeterminedtime setting for data output (e.g. kinet-ics monitoring in “Raw Fluorescence”)or leave the interval at 00:00:00 so thatyou receive a printout of data everytime the PRINT key is pressed (e.g.analysis of unknown samples using agiven method).

3. Run your Microsoft Windows program.Open Hyperterminal (located in theaccessories directory), give your settinga name and choose a symbol.

4. In the “Connect Using:” box, select“direct to” COM2: or (COM1:) as port(depending on your computer), andclick OK.

Printout Capability/Computer Connection

39

5. Using the selection boxes in the nextscreen, configure the options as 9600baud, 8-bit, No parity, 1 stop bit, FlowControl: None.

6. To receive data to a file, select Transferand capture text. Accepting default willput the file which you name in thec:\windows directory called capture.txtand the file will be an ASCII text file.

7. Operate the Quantech fluorometerusing either a “Method” or “Raw Flores-cence.” Pressing the PRINT key on thefluorometer will initiate the transfer ofdata to the computer. You should seethe data on the computer screen.

8. When you are finished operating theQuantech fluorometer, save your file.You can exit Hyperterminal or set up anew experiment.

9. Hyperterminal data can be manipulatedand graphed in Excel®.

Procomm

1. Connect the 9 pin serial cable from thefluorometer RS-232 port to COM2: portor (COM1: port) on back of computer.

2. * Access “Set Printer and Print Options”in the “Advanced Functions” menu (seeflowchart). You may set your printinterval to a predetermined time settingfor data output (e.g. kinetics monitoringin “Raw Fluorescence”) or leave theinterval at 00:00:00 so that you receive

PRINTOUT CAPABILITY/COMPUTER CONNECTION

40

a printout of data every time the PRINTkey is pressed (e.g. analysis of unknownsamples using a given method).

3. Open DOS window and change directoryto procomm directory.

4. Open procomm executable.

5. Press ALT-F10 for configuration screen.

6. Enter into Line Settings screen by typingALT-P.

7. Type in 11 <Enter>, sets parameters as9600, 8, N, 1.

8. Type in 21 <Enter>, sets to COM2: or(Type in 20 <Enter>, sets to COM1:),depending on your computer.

9. Type in 24 <Enter>, saves settings.

10. Type ESC to exit setup menu.

11. Procomm is now ready to accept inputfrom the Quantech fluorometer when thePRINT key on the unit is pressed.

12. To begin downloading a file, Type ALT-F1,and give the file an appropriate namewhen prompted.


41

The optional Epson™ printer (part no. AY1095X1)provides a paper tape record for future referenceand verifies fluorometer performance andfluorescence analysis.

During fluorescence analysis, the printer prints arecord of the method of analysis, sample number,concentration and fluorescence intensity units ata preset time interval.

Connecting and Starting thePrinter

1. Make sure the printer and Quantechfluorometer are turned OFF.

2. Connect the Quantech fluorometer tothe printer via the RS232 port on thefluorometer. Use the tan printer cableincluded with the Epson printer. Plugthe printer power cord into an electri-cal outlet.

3. Turn the Quantech fluorometer ON.

4. Turn the printer ON.

5. Access “Set Printer and Print Op-tions” in the “Advanced Functions”menu of the Quantech fluorometer(see flowchart located in the Appen-dix). You may set the print interval toa predetermined time setting for dataoutput (e.g. kinetics monitoring in“Raw Fluorescence) or leave theinterval at 00:00:000 so you receivea printout of data each time thePRINT key is pressed (e.g. analysisof unknown samples using a given“Method.”

NoteSee the Appendix of thismanual for printer setup instruc-tions.

Figure A: Printer

ON/OFF Button


42

6. Operate the Quantech fluorometerusing either a “Method” or “RawFluorescence.” Pressing thePRINT key on the fluorometer willinitiate the transfer of data to theprinter to provide a printout similarto the sample shown.

7. Printing can be terminated byexiting the “Method” or “RawFluorescence” analysis that youperformed.

Sample printout


43

The Quantech Digital Filter fluorometer requiresminimal preventive maintenance. It should be keptclean and away from dusty environments orenvironments with temperature and humidityextremes.

Excitation and Emission FiltersThree different types of filters may be used in yourTurner Quantech Digital Filter Fluorometer:

• Solid Glass Filters

• Gelatin Filters

• Interference Coated Filters

Gelatin and Interference CoatedFiltersGelatin and Interference Coated filters are mostsusceptible to environmental conditions (e.g.temperature above 50°C, humidity and improperstorage and handling. Care should be taken whencleaning, handling and storing all filters to preventdamage or scratching. Scratching of the filter mayresult in the transmittance of light of an undesiredwavelength to the photomultiplier tube. Carefullyinspect the filters for cracks, scratches or otherdamage. Replace filters if necessary.

Dust particles may be removed from Gelatin andInterference Coated filters by brushing filter surfacelightly with a clean, dry camel’s hair brush or bygently blowing clean, dry air across the surface.Glass filters are the most durable and may becleaned with lens paper, however, make sure thatthe filter surface are not scratched in the process.

NoteDisassembly of excitation andemission filters is not recom-mended. Damage of the filtersduring disassembly is likely andis not covered under warranty.

Maintenance

44

Sample Chamber & CuvetteHolder CleaningOpen the chamber cover and view the chamberfrom the top. Remove the Excitation andEmission Filters and store them in a clean, dryplace. Remove the two Phillips head screwsholding the cuvette holder in place. Carefullyremove the cuvette holder so as to avoidscratching the curved surface mirror ordamaging the spring clip on the holder.Carefully clean the cuvette holder with a waterdampened cotton swab and sample chamberwith a water dampened soft cloth. Allowcomponents to dry thoroughly beforereassembly.

NotePlease do not remove the topcase to expose the interiorcomponents of the units unlessspecifically authorized byBarnstead/Thermolyne. Thereare few user serviceable parts inthe Quantech fluorometer.

MAINTENANCE

45

Base Model Exploded View

Exploded View

PC1095X2

TNX107

CEX183

PC1095X3

LM1095X1

TV1095X1

FA1095X1

46

(See Exploded View.)

Part No. Description

CEX183 Power Entry Module

FZX58 Fuse Holder, Drawer

FA1095X1 Fan Assembly

LM1095X1 Quartz-Halogen lamp assembly

ME1095X1 2 x 24 LCD assembly

PC1095X2 Control PCB

PC1095X3 Power Supply PCB

SWX172 Keypad PCB and cover

TNX107 Transformer

TV1095X1 Photomultiplier tube, 931B

TV1095X2 Photomultiplier tube, R928B IR & WB

FZX35 Fuses: 5 x 20, 500MA, 250V (2

required), 100V & 120V

FZX51 Fuses: 5x20,.315A, 250V (2 required)

Gaining Access to ReplacementParts

Opening the Top CaseIn order to gain access to most of the replacementparts listed, the top case of the unit must beopened.

Replacement Parts

CautionService must only be attemptedby trained and qualifiedpersonnel.

47

1. Make sure the unit is OFF and the powercord is unplugged from the electricaloutlet and the unit power entry module.

2. Using a Phillips head screwdriver, removethe four screws located inside the samplechamber. These four screws secure thetop case to the sample chamber/filterblock (see Figure 2).

3. Slide the unit forward off the edge of thebench or tabletop surface on which it isresting just enough to allow you to seetwo silver Phillips head screws on thebottom of the unit. One screw is locatedon the bottom front of the unit and theother is located by the left front “foot”(black rubber vibration suppression pad)of the unit. Remove these two screws.The top case of the instrument will now liftup and back via the rear panel hinge.

4. Reverse the order of the above steps toreplace and secure the top case of theinstrument.

REPLACEMENT PARTS

48

Theory of FluorescenceWhen a molecule absorbs radiation, an electronis promoted from one energy level in the moleculeto a higher energy level within the molecule,absorbing the energy. The molecule may releasethe absorbed energy and return the electron to itsground state by converting this energy tovibrational energy or releasing the energy in theform of light emitted by the molecule. If part of thisenergy is converted to vibrational energy thereminder (if radiated within 10

-8 seconds), is

emitted as light of lower energy (longerwavelength) than the absorbed energy. Thisproperty is called fluorescence. Atoms ormolecules which fluoresce have well-definedexcitation and emission spectra which allow bothqualitative and quantitative analysis of thematerial.

The shape of the excitation spectrum is that of theabsorbance curve of the molecule and isindependent of the wavelength at whichfluorescence is measured. Each molecule alsohas a characteristic number called the quantumefficiency, which is the ratio of the total number ofemitted photons to the total number of absorbedphotons. A non-fluorescent molecule is onewhose quantum efficiency is zero or so close tozero that the fluorescence is not measurable.Both the quantum efficiency and the shape of theemission spectrum are independent of thewavelength of the exciting light. If the excitinglight used is of a wavelength which is differentfrom that of the absorption peak, a smallestportion of the light will be absorbed andproportionately less light will be emitted,illustrating the constancy of the quantumefficiency. However, the shape and location of theemission spectrum will not change.

Appendix

49

Fluorescence/ConcentrationRelationshipsFluorescence is related to and dependent onconcentration according to the following equation:

F=KIøCl

Where;

F = Fluorescence reading observed on theinstrument.

K = A constant which accounts for instrumentalfactors, pH, T, electronics.

I = The intensity of the exciting light at awavelength.

C = The concentration of the fluorescingmolecule.

l = The path length of the solution or solid beingmeasured.

ø = The quantum efficiency of the molecule.

An examination of this equation shows that for agiven molecule, at a constant concentration, theamount of fluorescence observed can be affectedby:

• I, the intensity of the exciting wavelength.

• D, the path length of the molecules.

As a practical matter, this means that the onlylimits on the sensitivity of fluorescencemeasurements are the electrical noise in theinstrument, competing radiations, physical

APPENDIX

50

limitations, i.e. sample volume available,maximum energy available and maximumacceptable cell size.

The accuracy of measurement is excellentbecause the technique is a direct measure ofthe radiant energy being made.

Considerations

Blank FluorescenceOne of the most common limits on sensitivity isset by blank fluorescence. Blank fluorescencearises from the reagents or other sourcesseparate from the compound of interest.Ideally, the fluorescence of the “blank” shouldbe 000, relative to the fluorescence of thematerial being analyzed.

ReagentsUsually, little difficulty will be encountered withcommercially available reagent-gradechemicals. This is particularly true inlaboratories utilizing procedures where themajority of assays are done with visible lightactivating the fluorescence of the sample.

If the samples are under ultraviolet excitation,or at a high instrument sensitivity, then high“reagent blank” fluorescence may beencountered. Even spectro-quality solventssometimes contain traces of impurities whichmay fluoresce under ultraviolet light. Simpledistillation in an all glass apparatus frequentlyremedies this problem.

APPENDIX

51

There are also several companies marketingfluorescent-grade reagents for those who must workat very low concentration.

A frequent source of contaminants are theplasticizers in some plastics and rubber. Fordispensing distilled water, an all polyethylene orglass system is recommended. Water standing inrubber tubing may develop high fluorescence frommoieties contained in the tubing duringmanufacture.

StandardsStandards should always be stored as concentratedsolutions, out of the light and at low temperature.Standard solutions of low concentrations maydegrade with time. Dilutions to workingconcentrations should always be prepared asrequired using appropriate diluent sincefluorescence is usually pH dependent.

GlasswareBorosilicate glassware is recommended for generalstorage of reagents. Test tube or flasks may becapped with parafilm for mixing and storage,however, care should be taken when using thismaterial. For work at very low levels offluorescence, it may be necessary to rinse allglassware with an appropriate solvent. See theOperation section.

TemperatureFluorescence is usually more temperature sensitivethan absorption. Although not all fluorescentmaterial exhibit marked sensitivity, there areinstances where as much as a 2% reduction in

APPENDIX

52

fluorescence is observed for every degreecentigrade temperature rise. Standards,reagent blanks and sample should be at thesame temperature prior to measurement offluorescence. Sometimes drifting of fluorometerreadings may be observed due to a change inthe temperature of the sample while in thesample chamber.

pHThe pH of the solution may affect thefluorescent species being tested. Protonationchanges the resonance structures of themolecule, the equilibrium of these species andconsequently, the concentration of thefluorescent molecule. Check your applicationscarefully for the possibility of this problem. ThepH of standard solutions may change over timedue to the acidic nature of storage containermaterials such as glass. Similarly, thefluorescent compound may decompose under avariety of pH ranges.

MiscellaneousSome materials are photosensitive; that is, theydecompose when exposed to wavelengths theyabsorb. This property can be determined byinserting a sample solution into the instrumentand observing its change in fluorescence overa period of time.

A few materials exhibit delayed fluorescence.This property can be verified by placing thesolution in the instrument and observing a risein the fluorescence reading after a period oftime.

APPENDIX

53

Cuvette Matching ProcedureCuvettes can affect results and are important tothe performance characteristics of the instrument.The cuvettes for any given determination shouldbe matched so that their fluorometric response isidentical in the instrument.

When using the Mercury Vapor Line Lamp for theUV excitation source below 340 nm, you mustuse quartz cuvettes.

Cuvettes may be matched as follows:1. Install the excitation and emission filters

for the appropriate analytical procedure.(Usually cuvettes are matched for usewith a specific analysis.)

2. Fill each cuvette to be checked with thefluorescent substance to be analyzed.

3. Set zero fluorescence with the blankreference solution cuvette and then ob-serve the fluorescence of each of theindividual cuvettes, which have beeninserted into the cuvette holder andaligned according to any orientationindicators.

4. Observe the fluorescence for each cu-vette and select those whose fluores-cence agree closely.

APPENDIX

NoteWhen using excitation wave-lengths below 340 nm, usequartz cuvettes.

NoteFor best results use quartzcuvettes for all your visible andultraviolet applications.

NoteWhen using round cuvettes,align them the same way in theinstrument every time. Varia-tions in glass thickness andproperties can affect your re-sults.

54

Excitation SourcesThe Quartz-Halogen lamp in the Base, IR andWide Band models of the Quantech DigitalFilter Fluorometer provides light in thewavelength range or 340-750 nm. The MercuryLine Lamp (UV Lamp) in the Wide Band modelis not a continuous lamp.

APPENDIX

Figure 5: Distribution of Mercury Line Lamp Emission Lines

55

FiltersThe filters in your Turner Quantech Digital FilterFluorometer are used to perform two functions.The first is to allow only light of a specificwavelength to pass into the sample cell andexcite a specific molecule. The excitation filtersare always Narrow Band (NB) filters and arespecific for the compound of interest. Theemission filters are generally Sharp-Cut (SC)filters that allow only emitted light above a specificwavelength to pass into the photomultiplier tube.Since the photomultiplier tube is sensitive to awide range of wavelengths, a filter allows the userto choose the wavelength range detected by thephotomultiplier tube to reduce background noiseand increase sensitivity.

SelectionFor nearly all fluorescent determinations, therequired filters are recommended by B/T. In theevent that a new procedure is being developed,the following considerations should be applied inselecting the proper filters:

1. Some compounds have the ability todischarge some of the energy obtainedwhen they absorb light by emitting light ofa longer wavelength. These compoundsare fluorescent. The efficiency of thisprocess may be anywhere from a fractionof a percent to almost 100 percent. Aportion of the absorbed energy is re-leased in heat.

2. The intensity of the light emitted is propor-tional to the amount of exciting lightabsorbed. At low concentrations, theemitted light may be considered propor-tional to the concentration. In practice, the

APPENDIX

56

meter reading of the fluorometer islinear with concentration of the fluores-cent molecule involved. The rangebetween the lowest detectable sampleand the point where significant non-linearity occurs is normally a factor of104 to 105 in concentration of the fluo-rescent molecule.

3. If a compound is fluorescent, a widerange of energy which it absorbscauses it to fluoresce. The spectrum ofthe emitted light is normally quite broadand its shape and peak are indepen-dent of the wavelength of the excitinglight. The only variation is in intensity;where less light (energy) is absorbed,less light will be emitted. Proportionalityof fluorescence and concentration aremaintained even when the exciting lightand measured fluorescence are farremoved from the peaks.

Narrow Bandpass Filter (NB)This filter is used when the excitation andemission wavelengths are close together.Typical bandwidths are in the 8 to 10 nm range.They require precision manufacturingtechniques and cost more than the other typesof filters. The filter consists of a miniatureinterferometer in conjunction with blockingglasses. For the filter designated NB390 therange of wavelength passed by the filter is 385-395 nm. These filters are constructed with athin coating of material and are susceptible toscratching. Extreme care should be taken whenhandling these filters. See the PreventiveMaintenance section for maintenance.

APPENDIX

57

Some Narrow Bandpass Filters are comprised ofspecial optical glasses or a combination of glassor gelatin materials, and have bandwidths of 50 to100 nm. Examples of these excitation filters arethe NB360 (bandpass 40 nm) and the NB430(bandpass 90 nm). These filters are particularlyuseful where high sensitivity is required, andlower resolution is not a factor in distinguishingbetween different fluorescent moieties present inthe solution. By allowing more light to pass to thesample, a greater number of the fluorescencemolecules are excited and more emission light isproduced per sample volume.

Narrow Band Filters can also be used in theemission side filter holder when specific emittedwavelengths need to be distinguished amongmany emitting fluorescence present in solution.

Sharp-Cut Filters (SC)This type of filter passes all light of a longerwavelength than a given value and blocks all lightof shorter wavelengths. The transition from zeroto full transmittance (approximately 80-90percent) usually takes place over a region ofabout 40 nm. The characteristic wavelength of aSharp-Cut Filter is defined as the wavelength atwhich the filter has a transmittance of 37 %. Withmost filter manufacturers, there is about a 10 nmtolerance in the location of the longestwavelengths of emitted light which can bemeasured.

APPENDIX

58

Photomultiplier Tube (PMT)The photomultiplier tube in the Base ModelQuantech Digital Filter Fluorometer is sensitiveto wavelengths of 300-650 nm but falls offrapidly above 650 nm. The photomultiplier tubein the IR and Wide Band Quantech Units arered-sensitive to wavelengths of 185-870 nm.

The PMT detector in all fluorescence measuresall the light reaching it from the cuvette. Byusing a NB or SC filter, certain wavelengths oflight are permitted to be passed to the PMT andsubsequently detected. Unwanted wavelengthsare not allowed to pass to the PMT, and do notaffect the fluorescence measurement. The NBand SC filters select a part of the PMTresponse curve to measure.

Compatibility of FiltersTwo filters are compatible and may be used asexcitation and emission filters if thewavelengths of the light they transmit do notoverlap significantly. If such an overlap ispresent, scattered light from an even slightlyturbid sample or from an optical defect in thecuvette will reach the photomultiplier and beregistered as fluorescence.

The steps involved in selecting the bestcombination of excitation and emission filtersfor a given compound are as follows:

1. Ideally the fluorescence spectra of thecompound will be available. Look forthe excitation and emission filtersrequired in Tables I and II. If the nameof the compound is known, the usermay consult the manufacturer of thefluorescent compound to obtain the

APPENDIX

NoteIt is important that the bandpasses of the excitation andemission filters do not overlapas this will affect the measure-ment of the fluorescence emis-sion by the PMT.

59

absorbance maxima for the compoundand the fluorescent spectrum maxima forthe compound. Direct the inquiries to thetechnical service department of thechemical manufacturer.

2. If the fluorescence spectra are not avail-able, then proceed as follows:

Refer to the transmittance curves for thisdiscussion.

a. Determine the maximum absorptionwavelength of the compound with aspectrometer. Choose a filter which isclosest to the peak from Table I and usethis for excitation.

b. Having chosen a tentative excitation filter,it requires but a few minutes to insert aseries of Sharp-Cut (SC) emission filterscompatible with the excitation filter, com-paring the ratio of sample reading toreagent blank. If the data obtained indi-cates a fairly sharp-peaked emissioncurve, the possibility of using a NarrowPass Emission filter may be considered.Usually there is little advantage to aNarrow-Pass Emission filter, though theyare occasionally of the value where theemission peak falls close to the excitationbandpass.

APPENDIX

Sharp Cut Filter Transmittance Curves

Bandpass Filter Transmittance Curves

60

Filter Selection Chart(See Table I, Table II and FilterTransmittance Curves)

All filters are mounted in filter holdersspecifically designed for simple wavelengthselection in the Quantech Digital FilterFluorometer. With the exception of theNB360(40 nm) and the NB430 (90 nm), allnarrow band filters designated by the centerwavelength in nanometers have a 10nmnominal bandpass curve. Sharp-Cut Filters(SC) are long pass filters. The number followingthe SC is wavelength, in nanometers, at whichthey exhibit a transmission of 37%. Theytypically have a transmission of less than 1% ata wavelength 10 nm less than the ratedwavelength. The NB excitation filters may beused as emission filters where wavelengths areclose together but sensitivity will be reduced.

APPENDIX

61

Table I - Quantech Excitation and Emission Filters

B/T PART # DESCRIPTION

Narrow Band (NB) Filters May Be Used As Excitation or Emission Filters

LE1095X1 NB254 FilterLE1095X3 NB297 FilterLE1095X4 NB313 FilterLE1095X30 NB360 FilterLE1095X6 NB365 FilterLE1095X7 NB390 FilterLE1095X8 NB405 FilterLE1095X9 NB420 FilterLE1095X31 NB430 FilterLE1095X10 NB440 FilterLE1095X11 NB450 FilterLE1095X12 NB460 FilterLE1095X13 NB490 FilterLE1095X14 NB520 FilterLE1095X15 NB540 FilterLE1095X16 NB590 FilterLE1095X17 NB680 Filter

Sharp Cut (SC) Filters Are Only Used As Emission Filters

LE1095X18 SC415 FilterLE1095X19 SC430 FilterLE1095X20 SC450 FilterLE1095X21 SC475 FilterLE1095X22 SC500 FilterLE1095X23 SC515 FilterLE1095X24 SC535 FilterLE1095X25 SC550 FilterLE1095X26 SC585 FilterLE1095X27 SC605 FilterLE1095X28 SC635 FilterLE1095X29 SC665 Filter

APPENDIX

62

Table II - Filter Requirements for Common Applications

Filters and Wavelengths

Application Method Primary SecondaryExcitation Emission

Fluorescein NB490 SC515Chlorophyll NB440 SC665Quinine Sulfate NB360 SC415Thiamine NB360 SC430Riboflavin NB440 SC535Tocopherols NB360 SC515Catecholamines NB405 SC515Rhodamine NB540 SC585Protoporphyrin NB405 SC585Niacin condensation NB360 NB440Niacin cyanogen bromide NB440 SC515Vitamin A NB360 NB490Vitamin B1 NB360 SC430Vitamin B2 NB440 SC535Vitamin B6 condensation NB360 NB440Vitamin B6 CN:pyridoxal NB360 NB440Vitamin B12 NB440 SC515Ascorbic Acid (Vitamin C) Deutsch NB360 SC440Vitamin E Kofler NB360 SC515Vitamin K Kofler NB360 SC430Vitamin K Jansson NB360 SC415DNA DABA NB405 SC515DNA thymine NB360 NB440DNA ethidium bromide NB360 SC585DNA Hoechst 33258 NB360 SC430 or SC415DNA Thiazole orange NB490 SC515DNA Oxazole yellow NB460 SC500DNA PicoGreen NB490 SC515Protein NanoOrange NB490 SC585Aflatoxin Methanol Extraction NB360 SC430Beta-Galactosidase NB360 NB460Histamine OPT NB360 SC450 or NB450RNA RiboGreen NB490 SC515ssDNA/Oligos OliGreen NB490 SC515

PicoGreen, NanoOrange, RiboGreen, and OliGreen are registered trademarks ofMolecular Probes, Inc.

APPENDIX

63

NB versus SC Filter UseThe previous discussions detail which filters touse for an individual application, however thereare times when the results of the fluorescentmeasurements are not linear, or are affected byundesired fluorescent moieties. In theseinstances using a Narrow Bandpass Filter (NB) inthe emission side of the filter holder may givemore accurate results.

In some instances more than one compound ispresent which can absorb the excitation light. Ifthe undesired molecule subsequently does notfluoresce, then the undesired molecule will haveno effect on the reading of the fluorometer. Ifhowever the undesired molecule does fluorescethen the proper choice of emission filter isimportant. The SC filter allows light of a specificwavelength and longer to be passed through tothe photomultiplier tube. In the case when thislight is the result of emissions from two or moredifferent compounds in the sample, then a choiceof using a Narrow Bandpass versus Sharp-Cutfilter must be made. This is where fluorometry hasa distinct advantage over absorptionspectroscopy.

If the fluorescence maximum of the desiredcompound is greater than the undesired moiety,choose a Sharp-Cut (SC) filter with a longerwavelength cutoff, above the fluorescencemaximum of the moiety. In the case where theundesired moiety fluoresces at a wavelengthlonger than the compound of interest, choose aNarrow Bandpass (NB) filter closest to thefluorescence maximum of the compound ofinterest.

APPENDIX

64

If the fluorescent wavelengths of twocompounds overlap, then careful choice of theexcitation bandpass filter can decrease thecontribution to the fluorescent light emitted bythe undesired moiety. Choose the excitationrange for the desired compound using an NBfilter at wavelength where there is significantabsorption of light by the desired compoundwhile there is little or no absorbance of light bythe undesired compound.

Compromises Between NB and SC

FiltersUse of an NB filter in the Excitation side of thefilter holder allows light of a specific wavelengthto excite a molecule. The intensity of the lightpassed through to the sample depends on thelight source and the bandpass filter. Choosingan NB filter with a larger bandpass, like for theNB360 and NB430 filters, will allow for a largerspectral distribution of light to pass to thesample and excite more molecules. This mayincrease sensitivity of the rest. Care should betaken when using these filters. Use of an NBfilter on the emission side of the filter holderpasses less of the fluorescent light produced bythe sample to the photomultiplier tube. Whilebeing more specific for a wavelength range, thesensitivity of the test may be reduced.

APPENDIX

65

Figure B: Control Panel

Figure C: Inserting the Paper Roll

Paper Feed Button

Power Light

Paper Out Light

Error Light

NoteA colored stripe along the edgeof the paper indicates that youare approaching the end of thetape. When the stripe appears,approximately 23 inches ofpaper remain, enough for 2-3cycles. Replace the tape beforeit is completely exhausted toensure a complete record ofyour analyses.

Printer Setup

Power SwitchLocated on the front of the printer (AY1095X1),this switch turns power to the printer ON andOFF. (See Figure A.) The green POWER light willilluminate when the printer is ON.

Paper Feed ButtonPress this button once to advance the tape oneline, or hold the button down to advance papercontinuously. When the paper roll nears the end,the red PAPER OUT light will illuminate.

If the red ERROR light illuminates, the printer isoff line. This could be a result of the print headbeing too hot, in which case the printer willresume printing once it cools down; or—the papermay be jammed in the printer. Turn the printerOFF before checking for jammed paper. Removethe paper jam and turn the printer back ON. If theprinter still will not print, unplug it and referservicing to qualified personnel.

Installing the Paper RollWhen the red PAPER OUT light illuminates, it istime to replace the paper roll. Replace the paperroll as follows:

1. Turn the printer ON and open the chambercover.

2. Make sure the edge of the paper roll isstraight.

3. Insert the paper roll into the printer. (SeeFigure C.)

APPENDIX

Figure A: Printer

ON/OFF Button

66

4. Insert the paper straight into the paperslot (see Figure D). The paper will feedautomatically.

5. Tear off a strip of paper and close thecover. (The red PAPER OUT light willbe OFF if paper roll installation issuccessful.)

Sample Printout

APPENDIX

Figure D: Inserting the Paper Into thePaper Slot

67

Quantech Accessories

Quantech Excitation and Emission Filters

B/T PART # DESCRIPTION

Narrow Band (NB) Filters May Be Used As Excitation or Emission Filters

LE1095X1 NB254 FilterLE1095X3 NB297 FilterLE1095X4 NB313 FilterLE1095X30 NB360 FilterLE1095X6 NB365 FilterLE1095X7 NB390 FilterLE1095X8 NB405 FilterLE1095X9 NB420 FilterLE1095X31 NB430 FilterLE1095X10 NB440 FilterLE1095X11 NB450 FilterLE1095X12 NB460 FilterLE1095X13 NB490 FilterLE1095X14 NB520 FilterLE1095X15 NB540 FilterLE1095X16 NB590 FilterLE1095X17 NB680 Filter

Sharp Cut (SC) Filters Are Only Used As Emission Filters

LE1095X18 SC415 FilterLE1095X19 SC430 FilterLE1095X20 SC450 FilterLE1095X21 SC475 FilterLE1095X22 SC500 FilterLE1095X23 SC515 FilterLE1095X24 SC535 FilterLE1095X25 SC550 Filter

APPENDIX

68

Sharp Cut (SC) Filters Cont.LE1095X26 SC585 Filter

LE1095X27 SC605 FilterLE1095X28 SC635 FilterLE1095X29 SC665 Filter

AY1095X1 Epson PrinterWHX18 Serial Cable for Communication w/ Computer

43F00-42 UV Suprasil Quartz, Square Cuvettes, 3 - 5 ml, matched pair11F06-07 Cuvette, 12 x 75 mm Round Borosilicate, 1-5 ml, pkg. of 1211F08-01 Cuvette, 12 x 75 mm Round Borosilicate, 1-5 ml, pkg. of 5

APPENDIX

69

Start

Intro

LanguageSelect

Initialize15 minutes

MainMenu

Choose MethodDNA

Choose MethodAflatoxin

Choose MethodHistamine

Choose MethodRhodamine

Choose Method<New Method>

AdvancedFunctions(see p. 3)

Raw Fluorescence

(see p. 7)

Standard(see p. 2)

Standard(see p. 2)

Standard(see p. 2)

Standard(see p. 2)

Standard(see p. 2)

BACK

ENTER ENTER ENTER ENTER ENTER

Last Used

timeout

AP

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Flow

chart

Flow

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70

AP

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Standard

Change name?

Proper Filtersin Unit?

Standard Curve From Memory?

Enter no. ofPoints (2-9)

EnterConcentration

- - - -

Units(see p. 7)

Insert KnownSample

Insert BlankSample

Coef. of Determination1.00

Insert Unkown Sample

Concentrationxxxx.xxx <units>

ABCAaBbCcDd

Filter SelectionHelp Guide

ENTER

Sample1n

ENTER

ENTER

NO, ENTER

YES, ENTER

NO, ENTER

YES, ENTER

ENTER

NO, ENTER

Notes:Up inserts letterDown removes letterENTER finishes selection

ZERO

ENTER

YES, ENTER

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AdvancedFunctions

Set Date andTime

Time and Date Set to:xx/xx/xxxx xx:xx:xx

Reset System toDefault Settings

ClearMemory,Reboot

Advanced(see p. 4)View Stored Data

ViewStored

(see p. 8)

Set Monthxx/xx/xxxx

Set Dayxx/xx/xxxx

Set Yearxx/xx/xxxx

Set Hourxx:xx:xx

Set Minutesxx:xx:xx

Set Secondsxx:xx:xx

ENTER

ENTER ENTER ENTER ENTER ENTERENTER

ENTERENTER

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APPENDIX

Set

Gai

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Set

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Set

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Set

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Turn

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73

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Note: BACK always steps back one step.MENU always returns to MAIN MENU.

PMT in CTS:Null = (xxxx) Value = (xxxx)

Diag Selection Menu<RAW PMT Hardware>

Diag SoftwareVx.xx.xx

DiagnosticMenu

(from p. 4)

AtoD Counts fro G = 1:Lamp_Detector = (xxxx)

AtoD Counts for:Anaolg Output = (xxxx)

AtoD Counts for G = 1:PGA w/REF Input = (xxxx)

AtoD Counts for G = 1:PGA w/GND Input = (xxxx)

AtoD Counts for G = 1:PGA w/PMT Lumin = (xxxx)

AtoD Counts for G=1PGA w/PMT Fluor = (xxxx)

AtoD Counts for:PMT PreAmp Out = (xxxx)

DIAGNOSTICS MENU<AtoD IN Out/Status>

Out/Status(see p. 6)

AtoD Counts for:System + 12V Pwr = (xxxx)

AtoD Counts for:System + 6V Pwr = (xxxx)

AtoD Counts for:Door_PDetector = (xxxx)

AtoD Counts for: RDAC = MHPMT HV Level = (xxxx)

<—Gain

<—Gain

<—Gain

<—Gain

<—Gain

<—PMTHV

ZERO

RIGHT RIGHT

LEFT

up

up

up

up

up

up

up

up

up

Notes:<— Change gain setting—> Increase RDAC null count by 10UP increase RDAC null count by 1DOWN decrease RDAC null count by 1ENTER save current null count to memory

Flow

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Output: Use ON and OFFHalogen Lamp is (OFF)

Output: Use ON and OFFHalogen Level is (LOW)

Output: Use ON and OFFUV Lamp is (OFF)

Output: Use ON and OFFInjector Outputis (LOW)

Status:Door is (CLOSED)

Date (NVRAM) Time08/05/98 8:00:00

Out/Status(from p. 5)

RIGHT

RIGHT

RIGHT

RIGHT

RIGHT

RIGHT

Flow

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75

Units

EnterConcentration

xxxx.xx <units>

Auto SetGain

InsertBlank Sample

InsertUnknownSample

pg/ml

ng/ml

ug/ml

ug/dl

ppt

ppb

ppm

<FIU>

Insert Blank Sample?

RAW FLUORESCENCE(from p. 1)

RawConcentrationxxx.xx <units>

Insert Sample

ng/ul

up

ENTER

ZERO

YES, ENTER

ENTER

ENTER

up

up

up

up

up

up

up

NO, ENTER

AP

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76

Aflatoxin

Gain Codexx

Concentration #1xxxx

Concentration #n

AtoD Reading #1xxxx

AtoD Reading #nxxxx

Blank A/D Readingxx

Rhodamine

Regression Slopex.xxx

RegressionIntercept

xx.xx

R Squared Termx.xxx

ViewStored

(from p. 3)down

down

down

down

down

downdown

down

down

down

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Ordering Procedures

Please refer to the Specification Plate for the complete model number, serial number, andseries number when requesting service, replacement parts or in any correspondenceconcerning this unit.

All parts listed herein may be ordered from the Barnstead Thermolyne dealer from whomyou purchased this unit or can be obtained promptly from the factory. When service orreplacement parts are needed we ask that you check first with your dealer. If the dealercannot handle your request, then contact our Customer Service Department at319-556-2241 or 800-553-0039.

Prior to returning any materials to Barnstead Thermolyne Corp., please contact ourCustomer Service Department for a “Return Goods Authorization” number (RGA). MaterialReturned without an RGA number will be returned.

80

Barnstead|Thermolyne Corporation warrants that if a product manufactured byBarnstead|Thermolyne and sold by it within the continental United States or Canadaproves to be defective in material or construction, it will provide you, without charge, for aperiod of ninety (90) days, the labor, and a period of two (2) years, the parts, necessary toremedy any such defect. Outside the continental United States and Canada, the warrantyprovides, for two (2) years, the parts necessary to remedy any such defect. The warrantyperiod shall commence either six (6) months following the date the product is sold byBarnstead|Thermolyne or on the date it is purchased by the original retail consumer,whichever date occurs first.

All warranty inspections and repairs must be performed by and parts obtained from anauthorized Barnstead|Thermolyne dealer or Barnstead|Thermolyne (at its owndiscretion). Heating elements, however, because of their susceptibility to overheating andcontamination, must be returned to our factory, and if, upon inspection, it is concluded thatfailure is not due to excessive high temperature or contamination, warranty replacement willbe provided by Barnstead|Thermolyne. The name of the authorizedBarnstead|Thermolyne dealer nearest you may be obtained by calling 1-800-446-6060(319-556-2241) or writing to:

Barnstead|ThermolyneP.O. Box 797

2555 Kerper BoulevardDubuque, IA 52004-0797

USAFAX: (319) 589-0516

E-MAIL ADDRESS: [email protected]

Barnstead|Thermolyne’s sole obligation with respect to its product shall be to repair or(at its own discretion) replace the product. Under no circumstances shall it be liable forincidental or consequential damage.

THE WARRANTY STATED HEREIN IS THE SOLE WARRANTY APPLICABLE TOBarnstead|Thermolyne PRODUCTS. Barnstead|Thermolyne EXPRESSLY DISCLAIMSANY AND ALL OTHER WARRANTIES, EXPRESSED OR IMPLIED, INCLUDINGWARRANTIES OF MERCHANTABILITY OR FITNESS FOR USE.

Two Year Limited Warranty

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