HARDWARE GUIDE - BioTek · BIO-TEK FL600™ FLUORESCENCE PLATE READER For Research & Development...

BIO-TEK FL600FLUORESCENCEPLATE READER

HARDWARE GUIDE

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HIGHLAND PARK, BOX 998, WINOOSKI, VT 05404-0998 USA.(800) 451-5172 • (802) 655-4040 • FAX: (802) 655-7941

BIO-TEK FL600 ™

FLUORESCENCEPLATE READER

For Research & Development Use Only

HARDWARE GUIDE

Part No. 6001000June, 1998Revision D

Bio-Tek FL600 iii

NoticesBIO-TEK INSTRUMENTS, INC.Highland Park, Box 998Winooski, Vermont, USA05404-0998

Customer Service & Sales: 888-451-5171 Outside U.S.: 802-655-4040Service: 800-24-BIOTKSales FAX: 802-655-7941Service FAX: 802-655-3399E-MAIL: [email protected] [email protected]: http://www.biotek.com

Lionheart Technologies, Europe N.V.Sneeuwbeslaan 33B-2610 WilrijkBelgiumTEL: (32) 3-8290455FAX: (32) 3-8254749

Copyright Copyright 1997, Bio-Tek Instruments, Incorporated. No part of thispublication may be reproduced, transmitted, transcribed, stored in a retrievalsystem, or translated into any language without the written permission of Bio-TekInstruments, Incorporated.

TrademarksBio-Tek® is a registered trademark, and FL600™ and KC4™ are trademarks ofBio-Tek Instruments, Inc.. Windows® and Windows 95 are trademarks of theMicrosoft Corporation.

Restrictions and LiabilitiesInformation in this document is subject to change, and does not represent acommitment by Bio-Tek Instruments, Inc. Changes made to the information inthis document will be incorporated in new editions of the publication. Noresponsibility is assumed by Bio-Tek for the use or reliability of software orequipment that is not supplied by Bio-Tek, or its affiliated dealers.

Bio-Tek Part No. 6001000

iv Bio-Tek FL600

Revision History

Revision Date Changes

A 4/97 Release to production

B 10/97 Added Stay Clear safety symbol. Corrected Electrical, Optical,and Performance Specifications sections. Added CE Markinformation. Deleted Appendix A.

C 3/98 Preface: Added warranty. Page 2-1: Modified instructions forSetting Up the FL600 Reader. Page 5-2: Changed electricalspecification for fuses. Page 5-11: Modified text and data onserial communications to reflect parity differences between KC4reader software and FL600 diagnostic software. Page 5-13: Onfilter table, modified use of excitation filters and added two newfilter wavelengths.

D 6/98 Preface: Changed phone numbers and e-mail addresses.Chapter 4: Changed air filter maintenance procedure. Addedlamp replacement procedure.

Bio-Tek FL600 v

Warranty

This Warranty is limited and applies only to new products, except for computer-basedsoftware which is covered under a separate Warranty Policy, manufactured byBio-Tek® Instruments, Inc. (“Bio-Tek”). Bio-Tek makes no warranty whatsoeverregarding the condition of used products.

Bio-Tek warrants the instrument (hereinafter collectively referred to as “Products” or“Product”) for a period of one (1) year from the original purchase date against defectivematerials or workmanship. This Warranty is limited to the original purchaser (the“Purchaser”) and cannot be assigned or transferred. All claims under this LimitedWarranty must be made in writing to Bio-Tek, Attention: Service Department.Purchaser must ship the Product to Bio-Tek, postage pre-paid. Bio-Tek shall eitherrepair or replace with new or like-new, at its option and without cost to the Purchaser,any Product which in Bio-Tek’s sole judgment is defective by reason of defects in thematerials or workmanship.

This Warranty is VOID if the Product has been damaged by accident or misuse, or hasbeen damaged by abuse or negligence in the operation or maintenance of the Product,including without limitation unsafe operation, operation by untrained personnel, andfailure to perform routine maintenance. This Warranty is VOID if the Product has beenrepaired or altered by persons not authorized by Bio-Tek, or if the Product has had theserial number altered, effaced, or removed. This Warranty is VOID if any of theProducts has not been connected, installed or adjusted strictly in accordance withwritten directions furnished by Bio-Tek. Batteries, fuses, lightbulbs, and other“consumable” items used in any of the Products are not covered by this Warranty.Software utilized in conjunction with any of the Products is not covered by the terms ofthis Warranty but may be covered under a separate Bio-Tek software warranty.

We will continue to stock parts for a maximum period of five (5) years after themanufacturer of any equipment has been discontinued. Parts shall include all materials,charts, instructions, diagrams, and accessories that were furnished with the standardmodels.

THIS WARRANTY CONTAINS THE ENTIRE OBLIGATION OF BIO-TEKINSTRUMENTS, INC., AND NO OTHER WARRANTIES, EXPRESSED, IMPLIED,OR STATUTORY ARE GIVEN. PURCHASER AGREES TO ASSUME ALLLIABILITY FOR ANY DAMAGES AND/OR BODILY INJURY OR DEATHWHICH MAY RESULT FROM THE USE OR MISUSE OF ANY EQUIPMENT ORINSTRUMENT BY THE PURCHASER, HIS EMPLOYEES, AGENTS, ORCUSTOMERS, OTHER THAN THE EXPRESS WARRANTY CONTAINEDHEREIN. WE SHALL NOT BE RESPONSIBLE FOR ANY DIRECT ORCONSEQUENTIAL DAMAGES OF ANY KIND. THIS WARRANTY SHALL NOTBE CHANGED OR MODIFIED IN ANY WAY WITHOUT THE EXPRESSWRITTEN PERMISSION OF AN OFFICER OF BIO-TEK INSTRUMENTS, INC.

vi Bio-Tek FL600

Warnings and Precautions

When operated in a safe environment, according to the instructions in this manual,there are no known hazards associated with the instrument. However, theoperator should be aware of certain situations which could result in serious injury.

Hazard Warnings

Warning! Power Rating. The instrument must be connected to a powerreceptacle that provides voltage and current within the specified rating for thesystem. Use of an incompatible power receptacle may produce electrical shockand fire hazards.

Warning! Electrical Grounding . Never use a two-prong plug adapter toconnect primary power to the instrument. Use of a two-prong adapter disconnectsthe utility ground, creating a severe shock hazard. Always connect the systempower cord directly to a three-prong receptacle with a functional ground.

Warning! Internal Voltage . Always turn off the power switch and unplug thepower cord before cleaning the outer surfaces of the instrument or changingfilters.

Warning! Liquids . Avoid spilling liquids on the instrument; fluid seepage intointernal components creates a potential shock hazard. Wipe up all spillsimmediately. Do not operate the instrument if internal components have beenexposed to fluid. If the microplate appears to have been exposed to fluid, thereading chamber may also have been exposed. To protect the incubator, removethe top cover and clean the interior.

Precautions

The following precautions are provided to help you avoid damaging the system:

Caution: Service. The system should be serviced by Bio-Tek-authorized servicepersonnel. Only qualified technical personnel should perform troubleshooting andservice procedures on internal components.

Caution: Replacement Fuses. Use replacement fuses with the required currentrating and specification. Improper fuses or short-circuiting the fuse holders maycause fire or damage the instrument.

Bio-Tek FL600 vii

Caution: Environmental Conditions. Do not expose the system to temperatureextremes. System performance may be adversely affected.

Caution: Filter Settings. The software filter settings must match the actualfilters used in the reader, in order to prevent possible damage to thephotomultiplier and/or degradation of data.

Safety Symbols

The following warning and informational symbols may be found in variouslocations on the instrument. Only qualified personnel who recognize shockhazards and are familiar with the safety precautions should use this instrument.Read the manual carefully before operating this instrument.

Direct currentCourant continueGleichstromCorriente continuaCorrente continua

Alternating currentCourant alternatifDreiphasen-WechselstromCorriente AtternaCorrente alternata

Both direct and alternating currentCourant continu et courant alternatifAllstrom (Gleich - und WechselstromCorriente continua y corriente alternaCorrente continua e corrente alternata

Earth ground terminalBorne de terreErde (Bettriebserde)Borne de TierraTerra (di funzionamento)

Protective conductor terminalBorne de terre de protectionSchutzleiteransclussBorne de Tierra de ProtecciónTerra di protezione

On (Supply)Marche (alimentation)Ein (Verbindung mit dem Netz)ConnectadoChiuso

viii Bio-Tek FL600

Off (Supply) Arrest (alimentation)Aus (Trennung vom Netz)DesconectadoAperto (sconnessione dalla rete di alimentazione)

Caution (Refer to accompanying documents)Attention (voir documents d'accompanement)Achtung siehe BegleitpapiereAtención (vease los documentos incluidos)Attenzione, consultare la doc annessa

Caution, risk of electric shockAttention, risque de choc electriqueGefährliche elektrische SpannungAtención, riesgo de sacudida eléctricaAlta tensione (in questo documentoAlta tensione non significa "tensione pericolosa"come definito in IEC 417)

Stay clearNe pas toucher svp.Bitte nicht berührenNo tocario por favorSi prega di non toccare

Bio-Tek FL600 ix

Contents

1. Introducing the Bio-Tek FL600

Hardware Requirements 1-3Software Requirements 1-4

2. Getting Started

Setting Up the FL600 Reader 2-1Installing KC4™ and FL600 Diagnostic Software 2-2

Installing KC4 2-2Installing the Hardware Key 2-2Installing the FL600 Diagnostic Software (Diagnostics Use)2-2

Checking the FL600 System 2-4Using the FL600 Diagnostic Software 2-4Filter Wavelength Locations 2-5Checking the COM Port 2-5

Running System Diagnostics 2-7What to Do if the Tests Fail 2-8

Exiting the FL600 Diagnostic Software 2-9

3. Replacing Filters and Adjusting Probe Height

Adding and Replacing Filters 3-1Changing the Filters Inside the Reader 3-2Setting the Filter Positions in KC4 Software 3-6

Utilizing the Filter Adapter System 3-7Removing the Filter from the Adapter 3-7Inserting the Replacement Filter into the Adapter 3-8

Adjusting Top Optical Probe Height 3-8FL600 Interior Configuration Diagrams 3-10

4. Bio-Tek FL600 Maintenance

Maintaining the Reader 4-1How to Maintain the Exterior 4-1How to Replace the Air Filter 4-1How to Replace the Lamp 4-3

x Bio-Tek FL600

5. Technical Information

Bio-Tek FL600 Fluorescence/Absorbance Plate Reader 5-1Physical Dimensions 5-1Operating Conditions 5-1Electrical Specifications 5-2Optical Specifications 5-2Scanning Mechanism 5-3Temperature Control 5-4Performance Specifications 5-4CE Mark Approval 5-8

Computer Specifications for FL600 Diagnostic Software 5-10FL600 Serial Communications 5-11Ordering Filters 5-12

Fluorescence Filters 5-12Absorbance Filters 5-14

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This chapter introduces the Bio-Tek FL600 FluorescencePlate Reader. You'll find information on hardware and

software requirements and optional equipmentsuggestions.

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Thank you for selecting the Bio-Tek FL600. The FL600 is a powerfulfluorescence and absorbance measurement system which measuresthrough the use of a photomultiplier tube housed inside of a light-impermeable detection compartment. The FL600 is capable of quantifyingboth soluble and cell-associated fluorescence. The system consists of:

• The Bio-Tek FL600 Fluorescence Plate Reader

• KC4™ reader control and data reduction software

• FL600 System Software for diagnostics

• Accessories Kit

• Serial Communications Cable (P/N 75034)

Both software packages are for Windows-based PCs and provides the userwith full command of the operation of the X-Y scanner. FL600’s precisionX-Y mechanism scans a microplate with a sensitive optical detectionsystem to quantify the fluorescence or absorbance in each well.

Introduction to the Bio-Tek FL6001-2

Virtually any microplate type from 6 to 384 wells can be used with theFL600. Users can select any microplate from the FL600’s built-in list ofindustry-standard plates, or they can define their own unique plateformat(s).

The FL600 is designed to meet international radio frequency emissionstandards, EMC immunity standards, and user safety standards.

The Bio-Tek FL600 should be installed in a well-ventilated area, free fromdust and fumes. Keep the system away from direct sunlight and local heatsources.

Introduction to the Bio-Tek FL600 1-3

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• IBM™ 486 PC or clone 33 MHz clock

• 640 KB base memory, extended to 8 MB

• Color graphics monitor (VGA)

• Color graphics adapter (VGA)

• 120 MB hard drive

• High-density floppy disk drive 3½"

• 1 serial (COM) and 1 parallel (LPT) port

• Mouse and keyboard ports

• Microsoft or compatible mouse

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• IBM 486 PC or clone 100 MHz clock

• 640 KB base memory, extended to 8 MB

• Color graphics monitor (VGA)

• Color graphics adapter (VGA)


• High-density floppy disk drive 3½"

• 1 serial (COM) and 1 parallel (LPT) port

• Mouse and keyboard ports

• Microsoft or compatible mouse

• Surge/spike protector

Introduction to the Bio-Tek FL6001-4

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• IBM 486 / 100 MHz PC or better compatible

• Memory: 640 KB base, extended to 16 MB minimum


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• Microsoft Windows compatible printer

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• FL600 System Software, version 2 or higher

• KC4™ reader control and data reduction software

• MS-DOS version 5.0 or later

• Microsoft Windows version 3.1 or later, or MicrosoftWindows 95 (with 16 MB memory)

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This chapter discusses how to set up the Bio-Tek FL600hardware, check the communication port and run the

diagnostic tests, install and configure the FL600 software,and exit the FL600 system software.

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CAUTION: It is recommended that two people lift the FL600, which weighs about 23kg (50.6 lb.). Care must be taken when moving the device or removing it from thepackaging. Hold the device firmly from the bottom of the case when lifting it.

Setting up the Bio-Tek FL600 Fluorescence Plate Reader is simply a matter ofconnecting the system to the computer via the computer’s serial port, installingthe software, and setting up communications parameters. One of the serial portsmost likely already has a mouse connected to it (the FL600 software, since itoperates under Windows, requires a mouse).

Step 1: Set the incubator power switch to the appropriate voltage asfollows:

110V for 100-120V˜ operation, 220V for 200-240V˜ operation

Step 2: Turn off the PC.

Step 3: To connect the FL600, first attach the 9-pin D-type cable into thecomputer’s serial port (COM1, COM2, COM3, or COM4).

Step 4: Plug the other end of the cable into the Serial Data Port located atthe rear of the FL600 Fluorescence Plate Reader.

Step 5: Connect the power cable to the back of the Plate Reader.

Step 6: Plug all power cords into grounded, electrical outlets (use of a surgeprotector is strongly recommended).

Getting Started2-2

The FL600 and all computer hardware must be electrically grounded andconnected to a viable surge protector to avoid dangerous shocks and toavoid destroying the electronic components through electrical spikes andsurges.

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The FL600 is supplied with two Windows™ software packages, KC4™and FL600 Diagnostic Software.

KC4 controls the reader, reduces the fluorescence and absorbance data,and furnishes the reports.

The FL600 Diagnostic software provides diagnostics to test the reader.Although it contains other functions, these are undocumented andunsupported.

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Please consult your KC4™ User’s Guide. Place disk 1 into the floppydrive. Run Setup.exe by choosing the Run option from the File Menu ofyour Windows program. This routine will guide you through theinstallation of KC4 onto your hard disk.

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To use KC4 to control the reader and print data, the hardware key needs tobe installed on the parallel port of your computer. Please consult yourKC4 Operations manual for further instruction.

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To install the FL600 software, follow these steps:

Step 1: Turn on your computer, start Windows, and activate Windows’Program Manager, if necessary.

Step 2: Insert the FL600 Setup diskette into the floppy drive.

Step 3: Select Run from the Windows’ File menu.

Step 4: Type drive: setup, (i.e., b: setup).

Getting Started 2-3

Setup will indicate to you that all FL600 diagnostic software files will beinstalled into the drive and directory C:\FL600. You will be given theopportunity to change either the drive or the directory (or both). Setupcreates a new group called Bio-Tek FL600. Note that this is different fromthe KC4™ group created by the KC4™ software.

NOTE: Be sure that TrueType Fonts are enabled in the Fonts Section of theWindows Control Panel. FL600 requires certain TrueType fonts whenprinting reports. For more information on activating TrueType Fonts,please check your Windows User’s Guide.

Getting Started2-4

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Before you begin to use the FL600 software, you should first check thatthe FL600 settings are correct.

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Step 1: To start the FL600 diagnostic software:

� From within Windows, double-click on the FL600 icon.

� Depending on your computer speed, the FL600 diagnosticsoftware should boot up from within 15 to 60 seconds.Longer than that might indicate that your computer systemneeds more RAM or clock speed.

� The FL600 Plate Layout screen will appear:

Figure 2-1: FL600 Plate Layout Screen

Getting Started 2-5

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Refer to Chapter 3 for instructions on determining filter locations. Recordthe filter locations to be entered into KC4.

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Next, turn on the FL600 Reader using the switch located on the right sideof the front panel and check the COM Port Setting.

NOTE: When the Reader has been switched on, a green light next the switchlights up and the plate transport moves out, ready to load a plate.

Step 1: Pull down the Instrument menu and select Communications . Adialog box appears showing you the currently selectedcommunications port, through which the software is attempting tocommunicate with the FL600 Reader:

Figure 2-3: Communications Port Settings Screen

Getting Started2-6

Step 2: Check to ensure that the correct com port is selected.

� You can test the COM port to see if it is connected andcommunicating correctly by clicking on Test. If the FL600is switched on and connected correctly, you will see thefollowing dialog box appear:

Figure 2-4: Plate Model Information Window

NOTE: You will see an error message if any of the following conditions exist: theFL600 is not correctly connected; it is not switched on; the incorrectcommunications port is selected; there is a cable problem. Check thatthe reader is switched on and correctly connected to the computer, thatyou have selected the correct communications port, and that the cable isoperational. Then close the message window and try the test again.

� Click on OK to close the dialog box. You will return to thecommunications port information dialog box.

� Click on OK to save the selected communications port, ifyou’ve changed it.

NOTE: If you select Cancel , the communications window will close withoutrecording any changes.

Getting Started 2-7

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The FL600 diagnostic software will automatically test the functionality ofthe Plate Reader, using the diagnostic tests.

Step 1: Pull down the Instrument menu and select Diagnostics . You willsee the following window.

Figure 2-5: Initial FL600 Diagnostics Screen

Step 2: To start diagnostic tests, click on the Start button. Informationdetailing the status of the reader appears in the window as each testis completed. (To close the window without starting the diagnostictests, click on Close or Cancel .)

Getting Started2-8

� Once the FL600 has completed its self-diagnostic tests,your screen should look similar to the one that follows:

Figure 2-6: FL600 Diagnostics Screen After Testing

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If any of the diagnostic tests should fail, click on the Log... button justbelow Cancel . A log showing exactly what the communications betweenthe computer and the reader was will display on the screen.

� Print the log by selecting the File menu and choosing Print .

� Enter the requested details on the fax cover sheet (whichprints automatically with the log) and fax both sheets toBio-Tek’s Technical Assistance Center at (802) 655-3399in the USA or +32-3-825-47-49 in Europe. TechnicalAssistance personnel will review the problem and instructyou on what to do next.

� To close the log, select the File menu and choose Close .

� Then, to close the diagnostics window and return to themain screen, click on the Cancel button on the Diagnosticswindow.

Getting Started 2-9

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Step 1: Select Exit from the File menu. You will be prompted to save anyunsaved work. Click on Yes to save the changes and exit, No toexit without saving changes, or Cancel to forget all about exiting.

Step 2: You will be prompted as to whether or not you want to close thereader door, thus parking the plate transport inside the reader.

Figure 3-11: The Exit Window

To exit the software and have the door automatically closed, click in theClose reader door check box and click on OK to exit the software andclose the door (make sure you have removed your plate). Click Cancel tocontinue without exiting.

NOTE: You should always opt to close the reader door when you are shuttingdown the system to protect the optics from dust. Alternatively, thecarrier can be withdrawn and the reader door closed by pressing thebutton located in the recess under the plate carrier.

NOTE: You can remove this software from your system (save the diskettes)and reload it if you suspect that you are having any problems with theinstrument.

Getting Started2-10

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This chapter discusses how to add and replace filters andadjust probe height for the FL600.

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Adding new filters or replacing existing filter sets is a two-step process:

• Changing the excitation and emission or absorption filters inthe filter wheels inside the reader.

• Changing the settings to reflect the filter change in the KC4™.

The settings in the system software must correspond exactly to thephysical location and specification of the filters on the filter wheels. Anerror in the system software specification for the filter could damage theinstrument and affect the FL600 warranty.

3-2 Bio-Tek FL600

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Step 1: Ensure that the plate carrier is parked inside the reader.

� Press the red button located in a recess under the platecarrier so that the plate carrier moves inside the reader andstops.

Park Button

Figure 3-1: Parking the Plate Transport Mechanism(Plate Carrier)

Step 2: Power down the FL600, and disconnect the reader’s power cord.

Never operate the instrument with the cover off. Operating the readerwithout a cover poses a serious shock hazard and may damage thedetection circuit.

Exposing the inside measurement chamber to ambient light for morethan 30 minutes may cause saturation of the measurement device.Should this occur, please wait 24 hours before reading.

Step 3: Remove the reader’s cover.

� Using a 4 mm Allen wrench (supplied in the AccessoriesKit), remove the five screws (Figure 3-2) that secure thereader’s cover to the base.

� Remove the top cover and set it aside.

Bio-Tek FL600 3-3

Figure 3-2: Removing the cover screws

Step 4: Remove the covers on both filter wheels.

� Locate the two filter assemblies at the rear of the reader,adjacent to the printed circuit board (Figure 3-3).

Excitation FilterWheel Cover

Emission FilterWheel Cover

Figure 3-3: Filter wheel locations

� Remove the filter wheel covers by pulling up on the headsof the plastic snap catches. (Figure 3-4)

3-4 Bio-Tek FL600

Figure 3-4: Removing the cover on the emission filter wheel

Step 5: Remove the filters you wish to change, being careful not to touchthe filter’s glass surface, or other optical components. Use extremecare to avoid dropping the filter into the interior of the instrument.

� Rotate the filter wheel until you reach the filter you want tochange.

� Using filter forceps (available in the Accessory Kit),remove the filter cell by pulling it straight up and out of thewheel.

� If removing absorbance filters, remove the holdercontaining the filter, not the filter itself.

Step 6: Ensure that the glass surfaces of the filters are free of dust andsmudges.

Step 7: Insert the new filter, and note its wavelength and position on thefilter wheel.

Fluorescence Filters:

� Align the new filter with the filter wheel, using theillustration below (Figure 3-5). The arrow on the filter cellshould point away from the center of the filter wheel.

� Insert the new fluorescence filter, and seat the filter byapplying gentle, equal pressure on the edges of the filterhousing.

Helpful hint: If the filter is marked 485/20X:• 485 is the center wavelength;

• 20 is the bandwidth at one-half maximum transmission(BWHM) of the excitation (X) filter.

Bio-Tek FL600 3-5

y An emission filter has an “M” designation.

• A dual-function Excitation/Emission filter has an“X/M“ designation.

Absorbance Filters:

� Insert the filter into the filter holder, and insert thisassembly into the filter wheel.

� Absorbance filters do not need to be aligned.

Step 8: Note the filter’s position on the wheel (the molded letters A, B, Care to the right of each filter), and the wavelength that is printed onthe top or side of each filter cell.

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Figure 3-5: Aligning the filter with the filter wheel

Step 9: Replace the covers on the filter wheels. Put any unused filters in a secure place.

� Secure the filter wheel covers by gently placing the coverover the access hole and then pressing down on the plasticsnap covers.

� Fit the instrument cover, and secure it using the five screws.

� Seal the unused filters in a plastic container, and store themin a cool, dry location.

Step 10: Re-connect the power cable, and power up the reader.

� The plate carrier will move to the plate load position.

3-6 Bio-Tek FL600

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The second and most important step in changing or adding filters to thereader is changing the KC4 software filter settings to accurately reflect thenew filters and their position on the wheel. The values for filters in theKC4 software must match the values of the filters in the reader to avoiddamaging the reader and invalidating data.

NOTE 1: Settings are stored on the PC, not the FL600. Remember to reprogramthe filter settings if you move the unit to another PC.

Bio-Tek FL600 3-7

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The FL600 comes equipped with a set of “500 series” fluorescence filters,which fit directly into the filter wheel. Should your application requireabsorbance filters of different wavelengths than those originally suppliedwith your instrument, you may order them from Bio-Tek.

Absorbance wavelengths are available only in the smaller “287 series.”These filters must be fitted into a filter adapter before insertion into thefilter wheel. The following instructions explain how to change the filter inthe adapter.

The “287 series” filter system involves the filter, the filter adapter, aretaining ring, and a removal-insertion tool. The tool is a cylindrical objectabout two inches tall, with nodes protruding from the top and bottom.

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Step 1: Remove the filter adapter from the filter wheel, as described in thesection entitled Changing Filters Inside the Reader.

Step 2: Place the adapter on a clean table. Be careful not to work too closeto the edge of the table, and give yourself plenty of space. (Thefilter is very small and easy to misplace when it pops out.)

Step 3: Place the adapter with the wide end face-down against the table.

Step 4: Insert the longer node of the tool into the top of the adapter.

Step 5: Place your thumbs on top of the tool and your other fingers alongthe edges underneath the adapter.

Step 6: Lift both objects as a unit, to approximately one inch off the table.Make sure that you keep the adapter parallel to the table.

Step 7: Firmly press the tool down into the adapter, until the filter andretaining ring pop out onto the table.

Step 8: Put the filter in a safe, clean place for future use.

3-8 Bio-Tek FL600

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Step 1: Ensure that the replacement filter is free of dust and smudges. Ifnot, carefully clean the filter with a cotton swab soaked in alcohol.

Step 2: Center the replacement filter on top of the retaining ring, and thenplace the adapter onto the filter, wide end down. If possible, alignthe window on the side of the adapter with the last four digits ofthe filter number, so that the four digits will appear in the window.

Step 3: Place both thumbs on either side of the top edge of the adapter, andpress down very firmly with equal pressure, until the assemblysnaps together.

Step 4: Insert the shorter node of the tool into the adapter, and press downfirmly to seat the filter.

Step 5: Install the filter adapter into the filter wheel, as described in thesection entitled Changing Filters Inside the Reader.

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Each FL600 model has been factory-configured with a top probe heightthat will provide maximum sensitivity for the specified plate thickness.Should you wish to change plate types, it may be necessary to reset theprobe height to prevent the probe from hitting the plate, or to maximizesensitivity.

To achieve maximal sensitivity when using the top optical probe it isnecessary to adjust the probe height as low as possible, while still allowingfree movement of the microplate underneath. Follow the directions belowto adjust the top probe height, referring to the diagrams in Figure 3-6.

Step 1: To withdraw the plate carrier into the instrument, press the platecarrier retrieval button located on the front underside of theinstrument. Shut off the instrument, and unplug the power cord.

Step 2: Using the 4 mm hex screw driver provided in the accessories box,remove the five machine screws holding down the top cover.These screws are located in the recessed pockets on the undersideedge of the bottom shroud. (See Figure 3-2.)

NOTE 1: Never operate the instrument with the cover off. Operatingthe reader without a cover poses a serious shock hazard and maydamage the detection circuit.

Bio-Tek FL600 3-9

NOTE 2: Exposing the inside measurement chamber to ambient lightfor more than 30 minutes may cause saturation of the measurementdevice. Should this occur, please wait 24 hours before reading.

Step 3: Reaching into the inside of the instrument, place a microplate onthe plate carrier, ensuring that it is the same type you usually use.

Step 4: Place the probe height adjustment shim (provided in theaccessories box) on the top of the plate, taking care to center it overthe plate. The end that contains the notch matching the top probesize must be oriented toward the optical probe.

Step 5: Using the 2.5 mm hex wrench provided, loosen the two set screwsholding the top probe in place. These two set screws are located onthe front surface of the top probe.

Step 6: Carefully raise the probe so that the bottom of the top probe isabove the top edge of the plate and shim on the plate carrier.

Step 7: Turn the X-axis and the Y-axis drive screws so that the appropriatenotch in the shim is located underneath the top probe.

Step 8: Carefully lower the top probe into the notch of the shim, takingcare not to damage the probe. Make sure that the probe is straightand vertical, and then tighten the set screws.

Step 9: Turn the X-axis drive screw so that the plate is no longer locatedunder the probe. Make sure that the plate carrier has not traveledso far as to open the door. If it has done so, back the plate up byturning the X-axis screw.

Step 10: Remove the microplate and shim from the plate carrier.

Step 11: Replace the top cover. Carefully align the five 4 mm screwsholding the top shroud in place, and then tighten them.

Step 12: Check the door for ease of movement. If the door movement isimpeded, try readjusting top shroud, making sure that it is onproperly and that the screws are tightened sufficiently.

Step 13: Attach the power cord, and turn on the power switch. The platecarrier should eject.

3-10 Bio-Tek FL600

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Figure 3-6: FL600 interior configuration

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This chapter explains the preventative maintenanceprocedures for the FL600, including how to

replace the air filter and the lamp.

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The FL600 requires regular, systematic maintenance to guarantee that itwill run in top form. This includes regularly cleaning the reader’s exterior,changing its air filter, and replacing its lamp.

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Exterior maintenance consists of simply wiping the reader with a dampcloth and mild detergent.

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Dust can affect flourescence values. The FL600 air filter should becleaned or replaced once every six months, more often if your site isespecially dusty.

Step 1: To access the air filter, snap the fan cover away from the right handunderside of the reader (on the same side as the plate transport), bygrabbing it between your thumb and forefinger and pullingdownward. (See Figure 4-1 and Figure 4-2.)

4-2 Bio-Tek FL600

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Figure 4-1

Step 2: Remove the air filter, a thin, grey piece of material.

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Figure 4-2

Step 3: Clean the filter by blowing the dust out with an air blower, and/orrinsing by hand with sudsy water and then clean water. Be sure todry the filter thoroughly before reinserting it into the fan cover.

Step 4: If the filter is damaged or difficult to clean, or if the fan cover isdamaged, replace the entire fan cover and filter with Bio-Tek P/N49840.

Step 5: Return the fan cover to its original position at the underside of thereader, and snap it back into place.

Bio-Tek FL600 4-3

Never use the FL600 without an air filter.

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NOTE: Full instructions for lamp replacement are provided with the lampreplacement kit.

CAUTION! Never touch the surface of the lamp, as any residue couldhamper illumination. We recommend that you wear gloves whentouching a new lamp.

Step 1: Make sure the plate transport mechanism is “parked.” To do so,press the red button in the recess under the plate carrier, so that theplate carrier moves inside the reader and stops.

park button

Figure 4-3

Step 2: Shut off the FL600, and then unplug both the power and computercables from the rear of the device.

4-4 Bio-Tek FL600

CAUTION! It is very dangerous to operate the FL600 with the coveroff, as there is a probability of electrical shock.

Step 3: Using the 4 mm Allen wrench provided in the accessories kit,unscrew the five screws holding the reader cover to the base.

Step 4: Remove the top cover, and set it aside.

Figure 4-4

� At the back of the FL600, on the left side, you will find thelamp assembly (as shown below).

Excitation Filter Wheel Emission Filter Wheel

LampAssembly

Figure 4-5

Step 5: Remove the two PMT wires from the cable harnesses on the lampcover, and disconnect the solenoid cable. (See Figure 4-6.)

Step 6: Unscrew the two thumb screws that hold the lamp cover onto thelamp assembly, and lift up the lamp cover. (See Figure 4-7.)

Bio-Tek FL600 4-5

Step 7: Using the 3mm Allen wrench provided in the accessories kit,unscrew the three screws mounting the lamp assembly to the lamphousing.

CAUTION! Fingerprints can ruin the FL600 quartz halogen lamp and render it unusable.

Step 8: Disconnect the lamp wires, and slide the lamp assembly away fromthe PMT base and toward the fan.

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Figure 4-6

Step 9: Carefully remove the lamp assembly from the lamp housing.

Step 10: Remove the lamp from the lamp assembly by removing the three3mm hex screws.

Step 11: Insert the new lamp (P/N 6000556S) into the lamp assembly, andthen replace the three screws.

Step 12: Remount the lamp assembly to the lamp housing, and screw downthe three 3mm hex screws. Be careful not to over-tighten them.

Step 13: Reconnect the two lamp wires in any configuration. Polarity is notan issue.

Step 14: Replace the lamp cover, making sure that cable attached to thesolenoid is routed through the center hole of the lamp cover.

4-6 Bio-Tek FL600

Step 15: Attach the lamp cover to the lamp assembly, using the two thumbscrews. Be careful not to over-tighten them.

Step 16: Reconnect the solenoid cable, and re-route the cables through thecable harness.

Step 17: Replace the reader cover, and attach it by screwing in the fivescrews.

Step 18: Plug in the power cable and the computer cable.

Step 19: Turn on the FL600. The plate transport mechanism willautomatically move into the load position.

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This chapter contains a variety of technical information, includinghardware specifications for the Bio-Tek FL600 Fluorescence Plate

Reader.

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Height 247 mm (97 in.)

Width 707 mm (278 in.)

Depth 426 mm (168 in.)

Weight 23 kg (50.6 lb.)

Shipping Weight 30 kg (66 lb.)

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Temperature +4°C to +35°C

Relative Humidity 20% to 80%, non-condensing

5-2 Technical Info rmation

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Fuse 5.0 amp, time-delay T, glass, 5 mm x 20 mm, 250 volt-type(Part Number 46055)

Input Voltage 90-132 volts AC180-240 volts AC47-63 Hz, 525 VA max

The power supply has a auto ranging power input.

Internal Voltages ±15 volt DC, +24 volt DC, +5 volt DC,-950 volt DC

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Lamp Type M32-Type, Tungsten-Halogen, 75 wattoptically stabilized lamp in fan-cooled assembly

Lamp Life 2000 hours minimum

Excitation Range 300 nm - 650 nm filtersfitted in a motor driven wheel assembly

Emission Range 350 nm - 750 nm filtersfitted in a motor driven wheel assembly (completewith photomultiplier)

Absorbance Range 340 nm - 750 nm filters

Filter Wheel 6 positions

Detector Photomultiplier Tube

Probe Diameter The fully randomized Quartz fiber bundles areavailable in four sizes to meet user needs. Differentprobes can be fitted to the upper and lower positionsat the factory.

5.0 mm general purpose probe for all media up to96 wells.

3.0 mm general-purpose probe suitable for media upto 96 wells

Technical Information 5-3

1.5 mm special probe for 384-well plates withminimum crosstalk, reduced sensitivity on otherplates

1.0 mm special probe for high-density media.

Probe Selector Probe diameters can be different in the top read andbottom read positions.

Selection of top or bottom read position is under softwarecontrol.

The height of the top reading probe may need to be set bythe user for different plate heights. See the sectionentitled Adjusting Top Optical Probe Height in Chapter3.

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Speed 65 mm/second maximum

Resolution 5 microns

Repeatability within ±25 microns

X Drive 10.0 mm leadscrew

Y Drives 10.0 mm leadscrew

There are two (Y axis) Probe scanning mechanisms fitted for top and bottomread configurations

Probe Configurations Selection of top or bottom reader or optical density modeunder computer control


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The FL600 is fitted with a heater block which can be programmed to heat theplate to a temperature above ambient.

Temperature Range Ambient +6° Celsius to 50° Celsius

Uniformity Within ±0.5° Celsius across a 96 well Dynatech black platemeasured with thermal probes in the center and four cornerwells and 150 µl of Johnsons Baby Oil per well.

Time to Stabilize Within 30 minutes after switching on

Time to Cool Approx. 2.5 hours from 37° Celsius to 6° Celsiusabove the on-board ambient temperature referencesensor.

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Standard Plate Types All 96-well microplates and most 48- to 6-welltissue culture plates

High-Density Plate Types 384-well microplates.

Note: Reader performance will be optimized by factory-fitting of a smaller fibre optic probe 1.0mm or 1.5mmdiameter).

60, 72 and 96 well Terasaki platesNote: Optimum probe size is 1.0mm or 1.5mm diameter.A custom plate adapter is required.

Plate Sizes Minimum 126 mm × 84 mm

Maximum 133 mm × 90 mm


Sensitivity (Specified)

Bottom Reading 5.0mm Probe

10 pg/ml solution of Sodium Fluoroscein, in PBS,150 µl per well Signal to Noise Ratio of 2 or greater.

Excitation 485/20, Emission 530/25Hellma 96-well quartz plate.

125 ng/ml solution of Propidium Iodide, in PBS,150 µl per well Signal to Noise Ratio of 2 or greater.

Excitation 485/20, Emission 645/40Cestar 3603 96-well plate.

Top Reading 5.0mm Probe

20 pg/ml solution of Sodium Fluoroscein, in PBS,300 µl per well Signal to Noise Ratio of 2 or greater.

Excitation 485/20, Emission 530/25Dynatech 96-well black plate.

160 pg/ml solution of Methylumbelliferone, in CBB,300 µl per well Signal to Noise Ratio of 2 or greater.Excitation 360/40, Emission 460/40Dynatech 96-well black plate.

125 ng/ml solution of Propidium Iodide, in PBS,300 µl per well Signal to Noise Ratio of 2 or greater.Excitation 485/20, Emission 645/40Dynatech 96-well black plate.

Sensitivity (Typical) Typical sensitivities can be two to three timesmore sensitive than specified


Data Capture Up to 6 different filter sets and sensitivities perscan; maximum of 300 scans (1800 repeat readsof the plate); wait time of up to 9999 secondsbetween each read of a plate

Kinetic Speed 30 seconds/plate, using Rapid Mode

4-6 seconds/column (depending on computerspeed), using Rapid Mode

5-10 reads/second/filter set, using Fast Kineticmode

1 read/3 seconds/2 filter sets, using Fast Kineticmode

End Point Speed 48 seconds/plate including in and out time usingrapid mode, in fluorescence.

Static Mode The probe will remain stationary under a well 0.1,0.35, 0.5, or 2 seconds before taking a reading. Asingle sample can be taken or 2, 3, 4, 5, 10, 20, 50,or 100 readings that are averaged. Time betweensamples can range from 10 ms to 2550 ms.

Plate Shaker The plate may be shaken at three settings for aperiod of 1 to 255 seconds. The settings are:

High 25 Hz 0.6 mm travelMedium 17 Hz 0.9 mm travelLow 12.5 Hz 1.2 mm travel


Fixed References The instrument contains 4 embedded referenceswhich are optionally read at the start of each scan.The references are:

1. Black Acetal Reference Disk used for OpticalDensity 0% transmission and fluorescencebackground

2. Green Reference Disk for fluorescence withExcitation 485 and Emission 530nm.

3. Brown Reference Disk for fluorescence withExcitation 590 and Emission 645nm.

4. Clear Optical path used for Optical Density100% transmission


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The device has been type tested by an independent testing laboratory and foundto meet the requirements of EC Directive 89/336/EEC for Radiated Emissionsand Line Conducted Emissions. Verification was to the limits and methods ofEN 55022:1995. The device is classified as EN 55022, Class A.

E N 5 0 0 8 2 - 1 I M M U N I T Y

The device was also tested and found to meet requirements for ElectrostaticDischarge Susceptibility, Radiated Susceptibility, and Electrical FastTransient/Burst Susceptibility. Verification of compliance was conducted tothe limits and methods of EN 50082-1:1992, IEC 1000-4-2:1995, EN 61000-4-3:1996, and IEC 1000-4-4:1995.

U S A F C C C L A S S A

Warning: Changes or modifications to this unit not expressly approved by themanufacturer could void the user's authority to operate the equipment.

This equipment has been tested and found to comply with the limits for a ClassA digital device, pursuant to Part 15 of the FCC Rules.

These limits are designed to provide reasonable protection against harmfulinterference when the equipment is operated in a commercial environment.Like all similar equipment, this equipment generates, uses, and can radiateradio frequency energy and, if not installed and used in accordance with theinstruction manual, may cause harmful interference to radio communications.Operation of this equipment in a residential area could cause interference, inwhich case the user will be required to correct the interference at his ownexpense.


C A N A D I A N D E P A R T M E N T O F C O M M U N I C A T I O N S C L A S S A

This digital apparatus does not exceed Class A limits for radio emissions fromdigital apparatus set out in the Radio Interference Regulations of the CanadianDepartment of Communications.

Le present appareil numerique n'met pas du bruits radioelectriques depassantles limites applicables aux appareils numerique de la Class A prescrites dans leReglement sur le brouillage radioelectrique edicte par le ministere desCommunications du Canada.

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This device has been type tested by an independent laboratory and found tomeet the requirements of:

E C D I R E C T I V E 7 3 / 2 3 / E E C , L O W - V O L T A G E D I R E C T I V E

EN 61010-1: “Safety requirement for electrical equipment for measurement,control and laboratory use. Part 1 General requirements.”


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Base Unit IBM 486 PC or clone 33 MHz clock640 KB base memory, extended to 8 MBColor graphics monitor (VGA)Color graphics adapter (VGA)120 MB hard driveHigh-density floppy disk drive 3½"1 serial and 1 parallel portMicrosoft Windows compatible mouse and keyboard ports

Printer Microsoft Windows compatible printer

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Base Unit IBM 486 DX PC or clone 100 MHz clock or better

640 KB base memory, extended to 8 MBColor graphics monitor (VGA)Color graphics adapter (VGA)120 MB hard driveHigh-density floppy disk drive 3½"1 serial and 1 parallel portMicrosoft Windows compatible mouse and keyboard portsSurge/spike protector

Printer Microsoft Windows compatible printer

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Base Unit IBM 486 DX PC or clone 100 MHz clock or better640K base memory, extended to 16 MB minimum250 MB hard drive

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MSDOS version 5.0 or higherBio-Tek FL600 System software version 2Bio-Tek KC4 reader control and data reduction softwareMicrosoft Windows version 3.1 or higher

Bio-Tek FL600 System software is compatible with Windows 95 (16 MB of RAMis recommended)


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The reader communicates with the computer via a 9-pin D-type male-femaleconnector (P/N 75034).

The connector is wired as follows.

PIN Computer FL600

1 DCD Not used

2 RX TX data to computer

3 TX RX data fromcomputer

4 DTR Not used

5 GND Signal ground

6 DSR Not used

7 RTS Linked to pin 8

8 CTS Linked to pin 7

9 RI Not used

Serial PortParameter

KC4 ReaderControl and Data

ReductionSoftware

FL600DiagnosticSoftware

Baud rate 9600 9600

Data bits 8 8

Parity Even None

Stop bit 1 1

Note: The FL600 Diagnostic Software settings can not be changed- througheither software or hardware tools.


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The following list reflects standard and custom filters for use with the FL600. For thelatest additions to this list, please contact Bio-Tek’s Customer Service Department.

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These are high-quality interference filters fitted in 22 mm diameter cells.

NOTE: The positions of each filter in the excitation and emission wheels can be user-configured for minimum switch time of pairs, in kinetics applications using two-color fluorescence.

FL500 AND FL600 EXCITATION/EMISSION FILTERS

Wavelength/Bandwidth

PartNumber

FL500 use(std 420-

700+nmversions)

Comments

360/40 5002020 Excitation only Standard Filter

380/20 5002028 Excitation only


420/50 5002006 Both

440/30 5002027 Both

440/40 5002007 Both

450/50 5002008 Both

460/40 5002022 Both Standard Filter.

485/20 5002021 Both Standard Filter

485/40 5002009 Both

508/20 5002018 Both

530/25 5002023 Both Standard Filter, used in both wheels

545/40 5002010 Both


Wavelength/

Bandwidth

Part

Number

FL500 use

(std 420-700+nmversions)

Comments

560/15 5002015 Both Alamar Blue™ Excitation Filter

560/20 5002011 Both

560/40 5002012 Both



620/40 5002013 Both

630/32 5002014 Both

633/10 5002017 Both

635/10 5002019 Both


680/30 5002029 Both

Excitation Filters


410/10 5002001 Excitation only 400/30 filter is preferred in mostcases


310/20 5002039 N/A



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All absorbance filters are placed in the emission filter wheel and require an adapter(part #6002000). See the section entitled Utilizing the Filter Adapter System inChapter 3, for further information. For the latest additions, please contact Bio-Tek’s Customer Service Department.

FL600 ABSORBANCE FILTERS

Wavelength Part Number (2874 pluswavelength)

340 2874340

380 2874380

405 2874405

410 2874410

415 2874415

440 2874440

450 2874450

465 2874465

470 2874470

490 2874490

515 2874515

530 2874530

540 2874540

550 2874550

562 2874562

570 2874570


Wavelength Part Number (2874 plusWavelength)

580 2874580

590 2874590

600 2874600

610 2874610

620 2874620

630 2874630

640 2874640

650 2874650

660 2874660

720 2874720

750 2874750


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