SPECTRAmax® PLUS384 User Manual
Contents
1. ZA PRAGA aa Va Appendix A A A 2 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual m r 3 O Printers and Cables Compatible The SPECTRAmax PLUS can be used with an Epson compatible strip Printers printer Compatibility may require changing the dip switch or pin settings on the printer or insertion of an emulation cartridge Please check your printer s manual to assess compatibility based on this criterion some printers will not meet this requirement Please contact your local Molecular Devices representative for information regarding recommended printers Cables Molecular Devices recommends that you use high quality double shielded cables to connect the SPECTRAmax PLUS to other peripheral instruments such as a printer or computer Choose cables that meet the following requirements Printer Cable for Stand alone operation only Centronics parallel male 25D to Centronics Serial Interface Cable contact Molecular Devices for specific pin out requirements Macintosh Male DB8 to male DB8 IBM Compatible Male DB8 to Female DB9 custom cable made by Molecular Devices SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual A 3 Appendix A A A 4 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Appendix B Accessories Items Available for Use with the SPECTRAmax PLUS Appendix B A B 2 SPECTRAmax PLUS Microplate Spectropbotometer Operator2. ids Periodically you should clean the outside surfaces of the SPECTRAmax PLUS using a cloth or sponge that has been dampened with water Do not use abrasive cleaners If required clean the surfaces using a mild soap solution diluted with water or a glass cleaner and then wipe with a damp cloth or sponge to remove any residue Do not spray cleaner onto the instrument If needed clean the microplate drawer or cuvette chamber using a cloth or sponge that has been dampened with water Should fluids spill in the drawer area when the drawer is out or into the cuvette chamber when the door is open they will be directed to a tray at the bottom of the instrument from which they will exit to the bench or counter beneath the instrument Wipe up any spills immediately Clean only the exterior of the unit and the microplate drawer or inside the cuvette chamber if necessary Never clean the inside of the instrument Do not allow excess water or other fluids to drip inside the instrument The fan filter on the bottom of the instrument requires periodic cleaning The frequency of the cleaning depends on how dusty your particular lab is and could range from once a month to once every six months 1 Turn power to the instrument OFF and then remove the power cord and cables from the back of the instrument 2 Make sure no plate or cuvette is in the instrument Tape the cuvette door closed Turn the instrument over so that it rests flat on the be
3. 8 well strip 77 K nm min 16 well strip SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 17 Chapter 1 A Temperature Regulation Reading chamber Isothermal when temperature regulation is not enabled Range 4 C above ambient to 45 C when temperature regulation enabled The ambient temperature must be gt 20 C to achieve temperature regulation at 45 C Resolution 0 10 Accuracy 1 0 C for microplate and cuvette chamber Temperature uniformity at equilibrium 0 5 C at 37 C Chamber warm up time 15 30 minutes measured on air after initiation of temperature regulation Temperature regulation 4 sensors Drift 0 2 C regulated Temperature regulation diagnostics Temperature regulation system is continuously monitored and updated Evaporation Plate lid required to minimize evaporative cooling Recommended microplate Flat bottom microplates with isolated wells and lid Automix with SOFTmax PRO Plate mixing modes Selectable off once prior to any reading and once prior to and be tween kinetic readings Plate mixing duration Selectable 1 to 999 seconds three second default Compatibility Microplate Standard and half area 96 well flat bottomed microplates 0 3 mL 384 well flat bottomed microplates Polystyrene plates for wave lengths above 340 nm UV transparent plates above 220 nm quartz plates above 190 nm Cuvette Standard height 45 mm cells with 10 mm pathlength 12 5 mm x 12 5 mm outside
4. with minimum inside width of 4 mm typical for 3 mL volume cells Test Tubes 12 x 75 mm test tubes can be used in the cuvette chamber with the test tube cover 1 18 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Instrument Description Display Operating panel Self diagnosis Spill control Computer interface Printer interface Microplates supported Operating temperature Operating humidity Stor age temperature Size h x w x d Weight Power consumption Line voltage Line frequency SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual General Instrument 2 x 20 character backlit LCD 9 key membrane keypad Continuous on board diagnostics Drawer mechanism reading chamber assembly is protected from ac cidental spillage by drainage ports 8 pin DIN RS 232 serial double shielding required Parallel 25 pin to Centronics double shielding required All 96 well and strip well microplates including lids Environmental 15 to 40 C 0 to 70 non condensing 20 to 65 C Physical 8 6 in 22 cm x 22 8 in 58 cm x 15 in 38 cm 30 Ib 13 6 kg lt 250 watts 90 250 VAC auto ranging 50 60 Hz z Chapter 2 A 2 2 SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual F Installation Installation Warnings Installation Cautions Unpacking 1 Always make sure the power switch on the instrument is in the OFF position and remove the power cord fr
5. Wavelength selection Wavelength bandwidth Wavelength accuracy Wavelength repeatability Photometric range Photometric resolution Photometric accuracy linearity microplate 0 2 0 OD Photometric accuracy linearity cuvette 0 2 0 OD Photometric precision repeatability 0 2 0 OD Stray light Photometric stabilization Photometric drift Calibration Optical alignment Light source Average lamp lifetime Illumination Photodetectors Photometric Performance 190 1000 nm Monochromator tunable in 1 nm increments lt 2 0 nm FWHM full width half maximum 1 0 nm across wavelength range 0 2 nm across all optical channels 0 3 to 4 000 OD 0 001 OD 190 1000 nm lt 1 0 and 0 006 OD 90 1000 nm lt 1 0 and 0 005 OD 190 1000 nm lt 1 0 and 0 003 OD lt 0 05 at 230 nm Instantaneous None continuous referencing of monochromatic output Automatic before first kinetic read and before every endpoint reading None required Xenon flash lamp 5 watts 1 billion flashes Top down microplates horizontal cuvettes Silicon photodiode SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Instrument Description Read time endpoint Standard read Speed read Kinetic read intervals Read time endpoint Kinetic read intervals Photometric Analysis Modes Stand Alone Single wavelength Absorbance or Transmittance reading of the cuvette or test tube Using SOFTma
6. maintenance or installation operation A EZR Never perform any operation on the instrument in an environ ment where liquids or potentially damaging gases are present A EZENN Risk of electrical shock Refer servicing to qualified personnel ANOTA Use of organic solvents such as dichloromethane may cause harm to the optics in the SPECTRAmax PLUS Extreme caution is advised when using organic solvents Always use a plate lid and avoid placing a plate containing these materials in the reading chamber for pro longed periods of time Damage caused by the use of incompatible or aggressive solvents is NOT covered by the instrument warranty A Never touch any of the optic mirrors filters or cables or their housing or manifold The optics are extremely delicate and critical toe of the instrument SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 3 nam Chapter 4 S ANNO Do not touch or loosen any screws or parts other than those spe cifically designated in the instructions Doing so could cause misalign ment and possibly void warranty General Keep the drawer closed when the instrument is not in use The drawer can be Cleaning Cleaning the Fan Filter 4 4 opened by pressing the DRAWER button Always close the drawer immediately prior to switching the instrument off EQ Wear gloves during any cleaning procedure that could involve contact with either hazardous or biohazardous materials or flu
7. on the right side of the SPECTRAmax PLUS and slides in and out of the reading chamber Springs on two sides of the drawer automatically position and hold a microplate in the proper position The drawer remains in the reading chamber during read cycles Microplate drawer operation varies depending upon the incubator status To open the drawer press the key If the incubator is turned on the drawer will remain open for approximately ten seconds after which a beeping sound will alert you approximately two seconds before the drawer closes automat ically Do not obstruct the movement of the drawer If you must retrieve a plate after an error condition or power outage and the drawer will not open it is possible to open it manually see Chapter 4 Maintenance and Troubleshooting Microplates The SPECTRAmax PLUS can accommodate standard 96 and 384 well microplates strip wells and filter bottom microplates When reading at wave lengths below 340 nm special UV transparent disposable or quartz microplates allowing transmission of the deep UV spectra must be used Not all manufacturers microplates are the same with regard to design materials or configuration Temperature uniformity within the microplate may vary depending on the type of microplate used 1 8 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual z O Instrument Description The Cuvette Chamber Figure 1 5 Cuvette Chamber Located a
8. that the products sold under this agreement are delivered free of the rightful claim of any third party by way of patent infringement or the like If the Buyer furnishes specifications to the Seller the Buyer agrees to hold the Seller harmless against any claim which arises out of compliance with the specifications Any description of the products contained in this agreement is for the sole purpose of identifying them Any such description is not part of the basis of the bargain and does not constitute a warranty that the products shall conform to that description Any sample or model used in connection with this agreement is for illustrative purposes only is not part of the basis of the bargain and is not to be construed as a warranty that the products will conform to the sample or model No affirmation of fact or promise made by the Seller whether or not in this agreement shall con stitute a warranty that the products will conform to the affirmation or the promise For the name of the nearest authorized Molecular Devices Corporation service facility please contact Molecular Devices at one of the following telephone numbers 408 747 1700 800 635 5577 U S and California SPECTRA max Microplate Spectrophotometer 100 240V 4A 50 60 HZ 4 0A TUV Rheinland GP 2 Lines Fused unplug before servicing Protected by US Patents 4 968 148 and5 112 134 Vor Wartungsarbeiten Netzstecker ziehen Molecular Sun iyvalevGA Protected by Canadi
9. to the instru ment off and then on again Manually ii If the drawer still remains closed turn the power off and using your thumb nail locate the groove in the upper left side wall of the door Open the door and with your index finger pull the microplate drawer out of the instrument do not force the drawer and remove the microplate This action will not harm the instrument but should only be taken if the first two options have failed to open the drawer If you are still unable to open the drawer contact Molecular Devices Technical Support or your local representative for assistance SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 11 Chapter 4 A 4 12 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual z 7 O Maintenance and Troubleshooting SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 13 Chapter 4 A 4 14 SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual z 7 O Maintenance and Troubleshooting SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 15 Chapter 4 A 4 16 SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual z 7 O Maintenance and Troubleshooting SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 17 Chapter 4 A 4 18 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Appendix A Printers and Cables Compatible Printers ag
10. type are used for emphasis Examples Press carefully to engage Do not press down NON A note provides information that will help you properly execute an action or procedure AMOUNT Indicates an action or condition that could potentially damage the instrument or one of its components or could result in loss of data A EZIN Indicates a situation that could result in potential injury to a person working with the system ORI VAN Indicates a condition involving potentially infectious biolog ical agents requiring that proper handling precautions be taken Absorbance A The amount of light absorbed by a solution To measure absorbance accurately it is necessary to eliminate light scatter In the absence of turbidity absorbance optical density A log 14 1 I incident light I transmitted light In this manual we use the terms absorbance and optical density interchangeably Optical Density OD The amount of light passing through a sample to a detector relative to the total amount of light available Optical Density includes absorbance of the sample plus light scatter from turbidity Transmittance T The ratio of transmitted light to the incident light T h T 100 T SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual vii viii SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Chapter 1 Instrument Description a A Kiso i ae oe 1 3 General Overview
11. will not be shown as raw OD values but are used to calculate subsequent sample reads which are shown on the printout and display Computer Control The SPECTRAmax PLUS is equipped with an 8 pin DIN RS 232 serial port through which a computer can communicate with and control the instrument SOF Tmax PRO Molecular Devices SOFTmax PRO software is a highly integrated program that can be used to control and collect data from the SPECTRAmax PLUS SOFTmax PRO is easy to use yet is powerful and flexible and makes it possible to access the full capabilities of the SPECTRAmax PLUS SOF Tmax PRO allows you to Read microplates for endpoint kinetic or spectral scans Use up to six wavelengths for endpoint and kinetic readings Perform Absorbance or Transmittance readings in the 190 to 1000 nm range Extend kinetic run times up to 99 hours SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 13 Chapter 1 1 14 Select your own read intervals for kinetic runs Specify the duration for Automix before and between readings Read the whole plate or a subset of microplate strips Read cuvettes for endpoint kinetic or spectral scans Use up to six wavelengths for endpoint and kinetic readings 1 Perform spectral scans from 190 to 1000 nm Extend kinetic run time up to 99 hours Select your own read intervals for kinetic runs Read a reference sample to be used with a single cuvette or a seri
12. 0 3 to 4 000 OD 0 001 OD 190 1000 nm lt 1 0 and 0 006 OD 90 1000 nm lt 1 0 and 0 005 OD 190 1000 nm lt 1 0 and 0 003 OD lt 0 05 at 230 nm Instantaneous None continuous referencing of monochromatic output Automatic before first kinetic read and before every endpoint reading None required Xenon flash lamp 5 watts 1 billion flashes Top down microplates horizontal cuvettes Silicon photodiode SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Instrument Description Read time endpoint Standard read Speed read Kinetic read intervals Read time endpoint Kinetic read intervals Cuvette Normal scan Cuvette Speed scan Microplate Normal scan Microplate Speed scan Photometric Analysis Modes Stand Alone Single wavelength Absorbance or Transmittance reading of the cuvette or test tube Using SOFTmax PRO e Express data as Absorbance or Transmittance Single wavelength reading of microplate and or cuvette Multiple wavelength up to six reading of microplate and or cuvette Kinetic and kinetic graphics of microplate and or cuvette Spectral scan 190 1000 nm of microplate and or cuvette Measurement Time calibration off Microplate 96 wells in 9 seconds single wavelength 96 wells in 9 N seconds N wavelengths 384 wells in 29 seconds single wavelength 384 wells in 29 N seconds N wavelengths 96 wells in 5 se
13. 1 3 PathCheck ea 1 4 Component Description 1 5 Control Panel 1 5 Control Panel Display 1 7 The Microplate Drawer 1 8 The Cuvette Chamber 1 9 The Backs Panelis ieri serioena a iia 1 10 Functional Description 1 11 Stand Alone Operation 1 11 Wavelength Selection 1 11 Temperature Regulation 1 11 Data Collecion add ass 1 12 Printed Data Output 1 12 Computer Control 1 13 Specifications oes ob eo eto eto ER 1 16 Chapter 1 A 1 2 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual F Instrument Description Introduction General Overview The SPECTRAmax PLUS microplate spectrophotometer provides rapid and sensitive UV VIS measurements of a variety of analytes including specific proteins nucleic acids and other molecules across a wide range of concentra tions It measures the optical density OD of samples in both 96 and 384 well microplates or in a cuvette at a selected wavelength for a single point in time endpoint over a specified period of time kinetic or over a selected wavelength range spectral scan MONO Figure 1 1 The SPECTRAmax PLUS 84 System The on board microprocessor calculates
14. SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual s Molecular Devices Corporation Molecular Devices 1311 Orleans Drive Sunnyvale California 94089 Part 0112 0056 Rev A PH Molecular Devices Molecular Devices Corporation SPECTRAmax PLUS Operator s Manual Copyright O Copyright 2000 Molecular Devices Corporation All rights reserved No part of this publication may be repro duced transmitted transcribed stored in a retrieval system or translated into any language or computer language in any form or by any means electronic mechanical magnetic optical chemical manual or otherwise without the prior written permission of Molecular Devices Corporation 1311 Orleans Drive Sunnyvale California 94089 United States of America Patents The SPECTRAmax PLUS and methods are protected by the following patents U S Patents 4 968 148 and 5 112 134 Canadian Patent 1 305 334 Other U S and International patents pending Trademarks SPECTRAplate SPECTRAstrip PathCheck and Automix are trademarks and SPECTRAmax and SOFT max are registered trademarks of Molecular Devices Corporation Apple and Macintosh are registered trademarks of Apple Computer Incorporated All other company and product names are trademarks or registered trademarks of their respective owners Disclaimer Molecular Devices Corporation reserves the right to change its products and services at any time to incorporate
15. agnostic checks to ensure that it is functioning correctly Turn the printer on at the same time Set the Temperature If elevated temperature within the cuvette or microplate chamber is required you should turn on the incubator first allowing enough time for the temperature to reach the set point before performing a reading When you first turn the instru ment on up to 30 minutes may be required for the temperature within the chamber to reach the set point To enable the incubator press the incubator TEMP On Off key The LCD dis play will update to show the temperature set point and the current chamber tem perature To change the temperature set point press the up or down arrows above and below TEMP until the desired temperature set point is shown in the display The microplate cuvette chamber temperature will be maintained at the set point until you disable temperature control by touching the incubator TEMP On Off key again When the incubator is off the drawer will open and the temperature within the chamber will begin returning to ambient NOVA Should you turn the incubator back on after a momentary shutdown allow about ten minutes for the control algorithm to fully stabilize the chamber temperature Select the Wavelength Press the up or down arrow keys above and below the A to increment or decre ment the wavelength setting 1 nm increments SPECTRAmax PLUS Microplate Spectrophotometer Operators Man
16. an Patent 1 305 334 Devices Made in USA Other US and International patents pending SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual iii SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual F Contents Fieures ri ea aaa vi Conventions UsedinthisManual vii Glossary Of NAM cad dog IO aed A vii Chapter 1 Instrument Description 1 1 Intreduetions 2 205402 cdo E ae ERS BATA NAA oe 1 3 General OVERVIEW utes ot trae NS 1 3 PathCheck cae ade m WG 0 eee RU ae wae 1 4 Component Description 1 5 Control Panel Goes Aa o A 1 5 Control Panel Display 1 7 The Microplate Drawer 1 8 The Cuyette Chamber cosas 1 9 IheBack Panel AA BEA ea 1 10 Functional Description 1 11 Stand Alone Operation 1 11 Wavelength Selection 1 11 Temperature Regulation 1 11 Data NA ee Aa 1 12 Printed Data Output 1 12 Computer Control 22 Actuate eee bas WEG de JE 1 13 Gaia s a ea E CRI IN E rd 1 16 Chapter 2 Installation 2 1 In
17. an Toda aia A 3 e ati dd tis tun A 3 Appendix B Accessories ss 63 0400 u 20 pae da B 1 Items Available for Use with the SPECTRAmax PLUS B 3 EEE II ORE NE E e ers I 1 Figure 1 1 SPECTRAmax PLUS 1 3 Figure 1 2 Major Areas of the SPECTRAmax PLUS 1 5 Figure 1 3 Control Panel 1 5 Figure 1 4 Microplate Drawer 1 8 Figure 1 5 Cuvette Chamber 1 9 Figure 1 6 Test Tube Cover 1 9 Figure 1 7 Components on the Back Panel of the SPECTRAma PLUSE Ser 2162 1 10 Figure 1 8 Sample Printout from Thermal Printer 1 13 Figure 2 1 View of Rear Panel 2 4 Figure 4 1 Power Switch Fuse Box and Power Receptacle 4 5 Figure 4 2 Removing the Fuse Box ana as last 4 6 Figure 4 3 The Fuse Box and Holder with Fuses Removed 4 6 Table 4 1 ErrorCodes 4 8 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual F Conventions Used in tbis Manual Glossary of Terms The names of keys that appear on the SPECTRAmax PLUS control panel are shown in boxed Helvetica type Example Drawer Italic and boldface
18. and reports the absorbance or 96 Trans mittance for each well of a microplate Data from multiple wavelengths can be acquired during a single reading if desired and different calculations can be made based on this data using SOFTmax PRO software including the subtrac tion of blanks use of standard curves etc Typical applications include endpoint assays quantitation of cytoproliferation by MTT reduction colorimetric protein assays protein measurement at 280 nm DNA RNA measurement at 260 nm and endpoint ELISAs and kinetic measurements enzyme studies such as determination of the activity of enzymes released from cells and kinetic ELISAs All standard 96 and 384 well microplates strip wells and filter bottom micro plates can be used in the SPECTRAmax PLUS 4 When reading at wavelengths below 340 nm special UV transparent disposable or quartz microplates and cuvettes that allow transmission of the fzr UV spectra must be used With SOFTmax PRO the contents of the wells in a microplate can be mixed automatically by shaking before each read cycle making it possible to perform kinetic analysis of solid phase enzyme mediated reactions mixing is not critical for liquid phase reactions The temperature of the microplate chamber can also be regulated if desired from 4 C above ambient to 45 C SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 3 nam Chapter 1 EE An optional printer f
19. arranty service prior to bringing or shipping the product for servicing At the time of requesting warranty service you must present proof of purchase documentation which includes the date of purchase and Molecular Devices Corporation must have the enclosed Warranty Registration form completed signed and returned by you within ten 10 working days of the date of delivery ii SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual This warranty only covers defects arising under normal usage and does not cover malfunctions of failures resulting from misuse abuse neglect alteration modification or repairs by other than an authorized Molecular Devices Cor poration service facility Repair or replacement as provided under this warranty is the exclusive remedy of the purchaser the Buyer Molecular Devices Corporation the Seller shall not be liable for any incidental or consequential damages for breach of any express or implied warranty on this product except to the extent required by applicable law The Seller specifically excludes all express and implied warranties including without limitation any implied warranty that the products sold under this agreement are merchantable or are fit for any particular purpose except such warranties expressly identified as warranties and set forth in the Seller s current operating manual catalog or written guarantee covering such product The Seller also makes no warranty
20. ate the heating cycles This tech nique results in accurate precise control of the chamber temperature with a temperature variation of the air inside the chamber of less than 1 0 C The tem perature uniformity within the microplate or cuvette will depend upon the design materials and or configuration of that component Data Collection The SPECTRAmax PLUS stores only the most recent cuvette reading in a buffer memory ARNOA Data in the buffer memory is lost when power to the SPECTRAmax PLUS is turned off This applies even to short power outages Do not turn the instrument off while important data remains in the buffer memory Printed Data Output An optional printer designed to sit on top of the instrument can be purchased for use with the SPECTRAmax PLUS During stand alone operation results are automatically printed as soon as a cuvette has been read A new cuvette can be loaded into the SPECTRAmax PLUS while the results from the first reading are being printed SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual z O Instrument Description HOLECULAR DEVICES CORI CFECTRAwaH PLUS DFERATDR DATE TIE AMP WAVELENGTH 65 na CUMETTE TEMPERATURE SETPOINT OFF CUVETTE TEMPERATURE 25 5 REFERENCE PAREM SAFLI SAMPLE ARG A SAHFLE SAMPLE SARPLE Figure 1 8 Sample Printout from Thermal Printer If you have performed a REF reading the REF values
21. bator Selects the wavelength that will be used for reading the cuvette manually Pressing the up or down arrow key scrolls up or down through the available wavelengths starting at the previ ous setting Pressing the up A or down arrow once incre ments or decrements the wavelength shown in the display by 1 nm pressing and holding either arrow increments or decre ments the wavelength shown in the display by 10 nm until it is released If you increment the setting to the highest limit 1000 nm and continue pressing the up A arrow the dis play returns to the lowest possible setting 190 nm and begins incrementing from there The inverse is true for decrementing by pressing the down arrow Setting the wavelength in SOFTmax PRO and reading using the soft ware will override the manual setting The control panel will not show the wavelength selected through SOFTmax PRO SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Instrument Description REF A reading of buffer water or air taken in the cuvette that is used as Ig to calculate Absorbance or Transmittance If no reference reading is taken the SPECTRAmax PL 76994 will use the Ip values stored in the NVRAM non volatile memory of the instrument Initiates the sample reading of the cuvette READ T A Toggle switch used to display cuvette data as percent transmission or absorbance Opens or closes toggles the microplate d
22. ce the SPECTRAmax PLUS on a level surface away from direct sunlight dust drafis vibration and moisture 2 Turn the instrument around so that the back of the instrument is facing you as shown in Figure 2 1 Fuse Box Cover Power Switch Power Cord Receptacle P Printer Centronics Computer RS 232 Figure 2 1 View of Rear Panel 3 7 8 Insert the appropriate Macintosh or PC serial interface cable into the RS 232 serial port on the SPECTRAmax PLUS Connect the other end to the serial port of the computer Insert the female end of the power cord into the power receptacle at the rear of the SPECTRAmax PLUS Connect the male end to a grounded power outlet of the appropriate voltage Molecular Devices recommends that you use a surge protector between the power cord and the grounded power outlet If you are using a printer for reading cuvettes in stand alone mode locate the printer port 25 pin parallel on the rear panel of the SPECTRAmax PLUS Connect one end of the cable to this port and the connect the other Centronics end to the printer Load paper into the printer according to the manufacturers instructions and connect the printer s power cord to the power outlet Turn the SPECTRAmax PLUS around so that the control panel now faces you Be sure no cables run beneath the instrument Leave at least three inches between the back of the instrument and the nearest objects or surfaces to ensure pr
23. conds single wavelength 96 wells in 5 N seconds N wavelengths 384 wells in 16 seconds single wavelength 384 wells in 16 N seconds N wavelengths 96 wells 9 second minimum interval between readings single wavelength 1 column 2 second minimum interval between readings single wavelength 384 wells 29 second minimum interval between readings single wavelength Cuvette 1 second single wavelength 2 second minimum interval between readings single wavelength Scan Speed 45 K nm min K wavelength interval 130 K nm min 33 K nm min 8 well strip 21 K nm min 16 well strip 135 K nm min 8 well strip 77 K nm min 16 well strip SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 17 Chapter 1 Specifications Thermal specifications for microplates used in the SPECTRAmax PLUS apply to flat bottom microplates with isolated wells All other microplate specifications apply to standard 96 well polystyrene flat bottom microplates Performance spec ifications for cuvette readings apply only to aqueous solutions having solute molal concentrations less than 0 4 M When pathlength compensation is applied to microplate absorbance measurements agreement with cuvette absorbance mea surements for the same solution requires that the solution volume in the micro plate well be between 100 pL and 300 pL Technical specifications are subject to change without notice Wavelength range
24. cular Devices representative Error Code Classifications Not all error messages are listed in this manual T he errors are grouped in rela tionship to possible causes as follows Error Code Numbers Possible Causes 100 199 Errors possibly caused by unrecognized commands being sent from the computer to the instrument 200 299 Errors probably due to a main board failure or an error in the firmware code Most of these errors require the assistance of Technical Support 300 399 Instrument errors due to either a main board failure or other system failure Most of these errors require the assistance of Technical Support 400 499 Errors caused by a motor motion failure Most of these errors require the assis tance of Technical Support 500 599 Errors due to failure or improper initialization of the instruments non volatile memory NVRAM All of these errors require the assistance of Technical Sup port Some errors shown in boldface in this manual are considered fatal if they are detected during power up the instrument will abort the power up sequence and display FATAL ERROR on the LCD panel Check the following table to see if you can do something to prevent the fatal error e g closing the cuvette door during the power up sequence After correcting the problem leave the instrument on for about five minutes turn it off and then back on If you con tinue to get the fatal error message o
25. cuvette door Cuvette door open while doing a read open 111 invalid read settings 200 299 Firmware Errors 200 assert failed Firmware error 201 bad error number Firmware error 202 receive queue overflow Caused by external device sending too much data over serial port and ignoring flow control 203 serial port parity error Parity bit error detected with incoming serial data 204 serial port overrun error Caused by host computer sending too much data and ignoring the flow control signal 205 serial port framing error 4 8 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Maintenance and Troubleshooting Table 4 1 Error Codes Messages and Notes about tbe Errors ERROR CODE ERROR MESSAGE NOTES 206 cmd generated too Firmware error much output 207 fatal trap Instrument error Instrument locks up 208 RTOS error Firmware error 209 stack overflow Firmware error 210 unknown interrupt Firmware error 300 399 Hardware Errors 300 thermistor faulty Unable to read a reasonable thermistor value Thermistor faulty or disconnected Main board problem or ambient temperature out of range 301 safe temperature limit A temperature of over 50 C detected on one or more of the 4 exceeded thermistors Temperature will be shut off and remain off until a successful completion of power up reset 302 low
26. e Box and Holder with Fuses Removed 8 Insert new SLOWBLOW rated fuses into the fuse holder Either end of the fuse may be forward 9 Insert the fuse box into the opening in the instrument making sure that the fuses are on the right side toward the power receptacle Press the fuse box into place making sure the cover snaps closed 10 Reconnect the power cord to the instrument and to the wall outlet and reconnect other cables previously disconnected SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual F Maintenance and Troubleshooting Error Codes and Probable Causes If a problem occurs during operation that causes an unrecoverable error the instrument will stop and an error code number will be shown in the display on the front panel To correct the problem call your local Molecular Devices repre sentative for assistance Error Messages The LCD will display Fatal Error codes when a situation arises that requires attention Any reading in progress will stop Warning messages do not stop a read ing but are logged in the error buffer Warning messages indicate a situation that requires attention but is not sufficient to stop or prevent a reading Examples of situations that might cause warning messages are low memory entries being out of range or operations that could result in loss of data These messages are gener ally self explanatory For assistance regarding warning messages contact your local Mole
27. erial 1 10 Power cord 1 10 2 4 B 3 Power switch 1 5 1 10 3 3 Preparing for a Reading 3 3 Print 1 12 Printed Data Output 1 12 Printer 1 4 1 12 1 13 2 4 3 3 Printer port 1 10 2 4 R Read mode 1 13 3 4 Read the Microplate 3 4 Read the microplate 1 13 1 14 Reference reading 1 7 3 4 Remote control 1 13 S Sample cuvette reading 1 7 Select the Wavelength s 3 3 Serial port 1 10 Set point 1 6 Set point temperature 1 7 Set the Temperature 3 3 Setting Up for Stand Alone Use 2 4 SOFT max PRO 1 3 1 5 1 13 Software SOFT max PRO 1 13 Specifications 1 16 Spills 4 4 cleaning 4 4 Stand alone operation 1 11 2 4 T Technical service 4 3 Technical Support 4 3 Temperature 1 11 1 12 3 4 equilibration 1 12 3 3 incubator 1 12 3 3 regulation 1 11 set point 1 6 1 12 3 3 setting 3 3 stabilization 1 12 Temperature Regulation 1 11 The Back Panel 1 10 The Microplate Drawer 1 8 1 9 Turn the Instrument On 3 3 U Unpacking 2 3 W Warranty ii Wavelength 3 4 range 1 16 Selection 1 11 Wavelength Selection 1 11 Wavelength setting 1 6 1 2 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual
28. es of cuvettes Use PathCheck to normalize the absorbance readings in each microplate well to a 1 cm pathlength Design microplate templates and cuvette templates to simplify data reduction Identify groups of wells or cuvettes with labels of your choice Identify individual wells or cuvettes with unique names Blank the entire plate groups and or individual wells Save instrument settings template formats and data analysis parameters for both microplates and cuvettes as assay protocol files and recall them for later use Rapid instrument and analysis set up for repeated microplate or cuvette assays Uniform analysis for equivalent microplates and cuvettes Turn the incubator on or off to control the temperature in the microplate drawer and cuvette port Acquire data from the SPECTRAmax PLUS Save data files for in depth analysis at a later time Save multiple microplates and cuvettes with individual template and data analysis parameters in one or more experiments in a single data file Pre read microplates Analyze kinetic and spectrum data as it is collected Use the Automix function to shake the microplate at preset intervals thereby mixing the contents of each well highly recommended for ELISAs and other solid phase enzyme mediated reactions Display data on screen Raw values reduced number or raw values with reduced number Raw microplate data in a microplate format Ranged data as integers betwee
29. from a local hardware store Make sure fuses are rated SLOWBLOW U S 4 amp time delay metric 4 amp 5 X 20 mm time delay 1 change fuses follow the steps below 1 Switch power to the instrument off and then remove the power cord from the outlet and from the SPECTRAmax PLUS power cord receptacle 2 Remove the printer cable and computer cable if connected from the back of the SPECTRAmax PLUS 3 Turn the instrument around for easy access to the rear panel 4 On the left hand side of the rear panel viewed from the back is the power switch fuse box and power cord receptacle As shown in the figures below press to the right of the black plastic cover of the fuse box to release it Pull the fuse box cover away from the instrument The fuse box will begin to slide forward Fuse Box Cover Figure 4 1 Power Switch Fuse Box and Power Receptacle SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 5 Chapter 4 5 Continue gently pulling the fuse box forward until it is free of the instru ment Figure 4 2 Removing the Fuse Box 6 When removed the fuse assembly will appear as shown in Figure 4 3 The holder inside contains two fuses 7 It is possible that only one of the fuses may have blown Molecular Devices recommends that you replace both fuses however to ensure continued proper operation Pull both fuses out of the holder and discard them Figure 4 3 The Fus
30. is plugged in When opened it provides access to the fuse box containing two fuses that are required for operation Printer port double shielded 25 pin parallel for use in stand alone opera tion plug the 25 pin end of the cable into this port the other Centronics end attaches to the port on the printer Computer port double shielded 8 pin RS 232 serial for use with an external computer plug the 8 pin DIN serial cable connecter into this port the other end attaches to the serial modem port of the computer Labels provide information about the SPECTRAmax PL US38 such as line voltage rating cautionary information serial number etc Record the serial number shown on this label for use when contacting Molecular Devices Tech nical Services 1 10 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual F Instrument Description Functional Description Stand Alone Operation The paragraphs that follow describe functions that can be activated during stand alone operation When operating the SPECTRAmax PLUS as a stand alone system you can obtain a fixed wavelength reading of the cuvette The wavelength is chosen by pressing the up or down arrows beneath the display labeled A Temperature regu lation of the cuvette microplate chamber can be enabled and set using the keys on the left side of the control panel and actual temperature can be monitored in the display To obtain a reading of the cuvette pres
31. it was commanded to in a faulty reasonable time 317 monitor ADC faulty 400 499 Motion Errors 400 carriage motion error Carriage did not move to either of its photo interrupts in a rea sonable time or can t find its photo interrupt 401 filter wheel error Filter wheel did not move to its photo interrupt in a reasonable time or cant find its photo interrupt 402 grating error Grating did not move to its photo interrupt in a reasonable time or cant find its photo interrupt 403 stage error Stage did not move to its photo interrupt in a reasonable time or cant find its photo interrupt 500 599 NVRAM Errors 500 NVRAM CRC corrupt The CRC for the NVRAM data is corrupt 501 NVRAM Grating cal Grating calibration data is unreasonable data bad 502 NVRAM Cuvette air cal Cuvette air calibration data is unreasonable data error 503 NVRAM Plate air cal Plate air calibration data is unreasonable data error 504 NVRAM Carriage offset Carriage offset data is unreasonable error 505 NVRAM Stage offset Stage offset data is unreasonable error 506 NVRAM Battery Time to replace the NVRAM battery U3 4 10 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual z 7 O Maintenance and Troubleshooting Opening the If an error occurs while the drawer is closed and you need to open the drawer y P Drawer press the Drawer key If the drawer does not open turn power
32. lash until the set temperature is reached If the incubator is on the LCD will show the current temperature along with the temperature set point To change the set point to any setting from ambient 4 to 45 C press either the up or down arrow above or below TEMP Select the desired measurement wavelength by pressing the up or down arrow near A Scroll up or down through the list of wavelengths shown in the LCD using the up or down arrows until the desired measurement wavelength is high lighted Load the prepared cuvette into the chamber being sure that the clear sides are left and right when facing the instrument Press the REF or READ CUVETTE key SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Chapter 4 Maintenance and Troubleshooting Technical Support 4 3 Warnings and Cautions 4 3 General ars GEE A 4 4 CIEMNE ee vit ue d eae O AE 4 4 Changing the Fan Filter 4 4 Changing the Fuses 4 5 Error Codes and Probable Causes 4 7 Error Messages 4 7 Opening the Drawer Manually 4 11 z Chapter 4 A 4 2 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Maintenance and Troubleshooting Technical Support Warnings and Cautions This chapter lists maintenance procedures and tro
33. le Printers A 1 Appendix C Accessories B 1 B Back Panel 1 10 Bandwidth 1 16 C Cables 2 4 A 3 connecting 2 4 serial 1 10 Calibration 1 16 Chapter 1 Instrument Description 1 1 Chapter 3 Stand Alone Operation 3 1 Chapter 4 Maintenance 4 1 Cleaning 4 4 Component Description 1 5 Computer Control 1 13 Computer control 1 13 Connections 1 5 1 10 cables 2 4 A 3 printer 1 10 2 4 Control panel 1 5 Control panel display 1 7 Cuvette 1 9 Cuvette chamber 1 5 1 9 D Data collection 1 12 printed output 1 12 Data Collection 1 12 Drawer open manually 4 11 Drawer key 1 7 1 8 1 11 E Error Messages 4 7 Error messages 4 7 F Fan filter cleaning 4 4 Fan filter 4 4 Functional Description 1 11 Fuse 4 5 changing 4 5 Fuses accessing 1 10 I If 3 4 Incubator 1 8 1 11 3 3 Installation 2 3 Installation Cautions Warnings 2 3 K Key READ 3 4 Keys 1 6 Keys control panel 1 5 L Labels 1 10 LCD Liquid Crystal Display 1 6 LCD liquid crystal display 4 7 LCD liquid Crystal Display 3 4 LCD Display 1 6 M Microplate 1 3 96 well 1 8 filter bottom 1 8 strip wells 1 8 temperature uniformity Microplate drawer 1 5 1 7 closing 1 8 1 8 1 8 incubator status 1 8 Microplates 1 8 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 1 Index O Opening the Drawer Manually 4 11 Operation Overview 3 4 P Pathlength correction 1 4 Ports 2 4 printer 1 10 s
34. light Not enough light detected to make an accurate measurement If doing a cuvette read the cuvette door may be open 303 unable to cal dark cur Too much stray light detected on power up faulty or discon rent nected pre amp boards 304 signal level saturation During a cuvette read could be due to cuvette door being open 305 reference level saturation During a cuvette read could be due to cuvette door being open 306 plate air cal fail low Minimum signal reference ratio not met during air calibration light 307 cuv air ref fail 308 stray light Light leak in reading chamber or cuvette door open Could also be a faulty pre amp board 309 front panel not respond LCD front panel bad or disconnected ing 310 311 312 gain calibration failed Power up calibration and check of signal path gain is out of tol erance Could be due to bad or disconnected pre amp or exces sive stray light SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 9 Chapter 4 Table 4 1 Error Codes Messages and Notes about tbe Errors ERROR CODE ERROR MESSAGE NOTES 313 reference gain check fail Power up check of the Reference amplifier s gain out of toler ance Could be due to bad or disconnected pre amp board or excessive stray light 314 low lamp level warning 315 can t find zero order On power up grating motor could not find zero order home position 316 grating motor driver Grating motor didn t move to where
35. lone functionality of the SPECTRAmax PLUS including setting the temperature inside the micro plate drawer and cuvette chamber setting the wavelength at which to read the cuvette toggling between displayed values as absorbance or percent transmission SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 5 Chapter 1 1 6 and opening or closing the microplate drawer Pressing a control panel key acti vates the function LCD A 2 X 20 character liquid crystal display which shows the current instrument set tings Keys FEE Incubator Enables disables the incubator function Incubator Allows you to enter a set point to regulate the microplate and cuvette chamber temperature Pressing the up or down arrow key scrolls up or down starting at the previous temperature setting or the default of 37 0 C if no setting had been made Pressing the up A or down arrow once incre ments or decrements the temperature shown in the display by 0 1 C pressing and holding either arrow increments or decre ments the temperature shown in the display by 1 C until it is released If you increment the setting to the highest limit 45 C and continue to press the up A arrow the display will not change If you decrement the setting to the lowest limit 15 C and continue to press the down W arrow the display will not change AKSIEGON 1f the incubator is disabled pressing the Temp On Off key will enable the incu
36. n 0 and 9 in a microplate format SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 2 O Instrument Description Threshold data as being above below or between set limits in a microplate format Gray scale data in seven shades of gray corresponding to high and low limits in a microplate format Kinetic or spectrum plots of all 96 or 384 microplate wells Enlarge the display of individual well plots and overlay multiple well plots Multiple cuvette data as raw ranged threshold gray scale kinetic or spectral scans Perform data analysis using SOFTmax PRO features Calculate maximum kinetic rates on non linear data Assign plate group or sample blanks Perform pathlength correction based on 1 cm cuvette readings Customize data analysis for each group in the template Create graphs with multiple plots Pick from nine curve fitting routines Analyze unknown samples against a standard curve Analyze and compare data within a plate between plates and between exper iments Multiple print formats Print all or individual sections of the data file Define and print a report containing only selected sections Customize the order of data file sections Export data in tab delimited ASCII format for use with Excel or other database programs or as JCAMP DX format for use with spectral analysis software For a complete description of the features of SOFTmax PRO refer to the SOFTmax PRO U
37. n power up record the error message number and contact Molecular Devices Technical Support or your local representative for assistance SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 4 7 Chapter 4 O If the instrument is functioning normally when using SOFTmax PRO no errors should be in the buffer except error number 100 Table 4 1 Error Codes Messages and Notes about the Errors ERROR CODE ERROR MESSAGE NOTES 100 199 Operator Errors 100 command not found Command string not recognized 101 invalid argument Command Argument not recognized 102 too many arguments Too many arguments after command 103 not enough arguments Missing arguments 104 input line too long Too many characters in the input line 105 command invalid sys Instrument could not perform the give command because it was tem busy busy doing another task Example Request a wavelength while the monochromator is in motion 106 command invalid mea Instrument could not perform command because a measurement surement in progress was in progress 107 no data to transfer Inputting transfer when there s no data in the buffer 108 data buffer full To many data sets in the buffer Can be caused by setting up a long kinetic and disconnecting computer or SOFTmax PRO is preempted by another application 109 error buffer overflow More than 65 errors in the buffer clear the buffer 110 stray light
38. nch 3 Pop the black fan cover off the remove the filter 4 Clean the filter by blowing clean canned air through it or by rinsing it first with water and then with alcohol and allowing it to dry completely 5 Place the clean dry filter over the fan and replace the black cover 6 Turn the instrument back over Remove the tape from the cuvette door Reconnect the power cord and cables to the instrument SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 5 Maintenance and Troubleshooting Changing the Fuses Fuses burn out occasionally and must be replaced If the instrument does not seem to be getting power after switching it on the LCD shows no display first check to see whether the power cord is securely plugged in to a functioning power outlet and to the receptacle at the rear of the SPECTRAmax PLUS If power failed while the SPECTRAmax PLUS was already on check that the power cord is not loose or disconnected and that power to the power outlet is function ing properly If these checks fail to remedy the loss of power follow the steps listed below to replace the fuses Spare fuses two U S and two metric are shipped with the instrument The U S and metric fuses are identical except for physical size They may be taped to the back of the SPECTRAmax PLUS3 If you no longer have spare fuses you may obtain new ones from Molecular Devices part numbers 4601 0013 for U S 4601 0014 for metric or
39. om the back of the instrument prior to any instal lation or relocation of the instrument 2 Do not operate the instrument in an environment where potentially damag ing liquids or gases are present 1 Do not touch or loosen any screws or parts other than those specifically desig nated in the instructions Doing so might cause misalignment and will void the instrument warranty The SPECTRAmax PLUS is packed in a specially designed carton Please retain the carton and the packing materials If the unit should need to be returned for repair you must use the original packing materials and carton for shipping If the carton has been damaged in transit it is particularly important that you retain it for inspection by the carrier in case there has also been damage to the instrument AEAN The SPECTRAmax PLUS weighs approximately 30 lb 13 6 kg and should be lifted with care It is recommended that two per sons lift the instrument together taking the proper precautions to avoid injury After examining the carton place it on a flat surface in the upright position Open the top of the box and lift the SPECTRAmax PLUS along with the packing materials around the ends up and out of the shipping box Remove the packing material from both ends of the instrument and set the instrument down carefully SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 2 3 Chapter 2 Setting Up 1 for Stand Alone Use 2 4 Pla
40. oper ventilation and cooling Remove the tape from the cuvette door Turn on the power to the SPECTRAmax PLUS 4 wait for the microplate drawer to open and remove the tape and protective covering from the drawer subplate SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual Chapter 3 Stand Alone Operation Prepare for a Reading 3 3 Turn the Instrument and Printer On 3 3 Set the Temperature 3 3 Select the Wavelength 3 3 Read the Cuvette 3 4 Operation Overview 3 4 z Chapter 3 A 3 2 SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual Stand Alone Operation Prepare for a Reading This chapter contains operating information for the SPECTRAmax PLUS Micro plate Cuvette Spectrophotometer If you are an experienced user of this instrument you can turn to the Operation Overview on page 3 4 for a quick review of the operat ing steps Only temperature selection and single wavelength cuvette readings are available in stand alone mode through front panel control To achieve full use of this instrument connect it to a computer and use SOFTmax PRO software Turn the Instrument and Printer On The power switch for the SPECTRAmax PLUS is located on the back panel Press the rocker switch to the ON position The instrument will automatically perform di
41. or use with the cuvette chamber in a stand alone mode can be purchased as an accessory TM PathCheck As predicted by the Beer Lambert law of light absorption absorbance is propor tional to the distance that light travels through the sample the longer the path length the higher the absorbance Microplate readers use a vertical light path so the distance of the light through the sample depends on the volume This variable pathlength makes it difficult to per form extinction based assays and confusing to compare results between micro plate readers and spectrophotometers Horizontal The standard pathlength of a cuvette is the conventional basis for quantifying the unique absorptivity properties of compounds in solution orar Quantitative analyses can be performed on the basis Light Path of extinction coefficients without standard curves e g protein at 280 nm nucleic acid estimates at 260 nm and NADH based enzyme assays When using a cuvette the pathlength is known and is independent of sample volume so absorbance is C proportional to concentration uvette In the microplate pathlength is dependent on the liquid volume so absorbance is proportional to both Vertical the concentration and the pathlength of the sample Standard curves are often used to determine analyte concentrations in vertical beam photometry of knowns yet errors can still arise from pipetting the Vertical Light Path es pipertng ern samples and
42. rature to a setting lower than ambient Addition ally the highest setting 45 C can be achieved only if the ambient tem perature is gt 20 C You can change the temperature set point by pressing the up A or the down arrow keys above and below the word TEMP until the desired set point is shown in the display Typically the microplate chamber will reach 37 0 C in 15 to 30 minutes The microplate chamber temperature is maintained at the set point until you press the incubator TEMP On Off key again turning temperature regulation off The microplate drawer will open and the temperature within the chamber will begin returning to ambient SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 11 Chapter 1 NY Should you turn the incubator back on after a momentary shutdown allow about ten minutes after reaching temperature for the control algo rithm to fully stabilize the microplate chamber temperature Temperature regulation and control of the microplate cuvette chamber are achieved through electric heaters a fan efficient insulation and temperature sen sors The heaters are located in the chamber which is insulated to maintain the temperature set point The sensors also mounted inside the chamber measure the air temperature The temperature feedback closed loop control algorithms compare the measured air temperature inside the chamber against the tempera ture set point and use the difference to calcul
43. rawer Whether or not the drawer will remain open depends on the incubator setting If the incubator is off the drawer will remain open if the incubator is on the drawer will close after approximately 10 seconds to assist in maintaining temperature control within the microplate chamber If the SPECTRAmax PLUS is being controlled by the computer software the READ CUVETTE and REF button will be inactive If the SPECTRAmax PLUS is idle all keys on the control panel are active During a kinetic run if the read interval is gt 30 seconds pressing the button will open the drawer The instrument will automat ically close the microplate drawer for the next reading Control Panel Display The left side of the display shows the temperature both actual and set point and whether or not the temperature is at the set point enunciator blinks if not at set point The middle of the display shows the wavelength The right side of the dis play shows the data received from the reading as either absorbance or percent transmission and indicates whether or not a reference measurement was made enunciator blinks if no reference reading was taken To change the contrast on the control panel press T A and the Temp up or down keys SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 7 Chapter 1 A The Microplate Drawer Al Indicator Figure 1 4 Microplate Drawer The microplate drawer is located
44. s Manual CO Accessories Items Available for Use witb tbe SPECTRAmax PLUS Part Number Cable RS 232 Macintosh to SPECTRAmax PLUS3 DIN HL DIN 9000 0091 Cable RS 232 PC to SPECTRAmax PLUS Opin 0 8 pin DIN du SA A a ees 9000 0149 Cable parallel printer 25 pin to Centronics 9000 0003 Cuvette Calibration Kit 9000 0161 Fuse 4 amp Time Delay 4601 0013 Fuse 4 amp 5 X 20 mm Time Delay 4601 0014 Power Cord US Canada Japan Mexico India 4400 0002 Power Cord Germany France Scandinavia Italy Korea 4400 0036 Power Cord UK Indonesia Singapore Malaysia 4400 0037 Power Cord Australia Hong Kong China 4400 0038 Printer a E T 9000 0189 SPECTRAplate Disposable UV transparent disposable microplates case of 50 R9013 SPECTRAplate Quartz UV transparent quartz microplate R8024 SPECTRAmar Mouse Pad ve ao pet e ai e taia aa en 9000 0133 SPECTRAtest PLUS Validation Package 0200 2405 Ti tOVEL e aco ce CO DC A DO RE A 2300 0277 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual B 3 Appendix B A B 4 SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual F Index A Appendix B Compatib
45. s either the REF key for a reference reading or the READ CUVETTE key for a sample reading The resulting data can be displayed as absorbance or percent transmission by toggling the T A key You can also open or close the drawer using the Drawer key on the right of the control panel Keep cuvette door closed when reading a cuvette Wavelength Selection The display on the control panel shows the currently selected measurement wave length A You can change the wavelength by pressing the up or down arrows until the display shows the desired wavelength The wavelength will wrap around to the top or bottom when the last selectable wavelength is reached Temperature Regulation The SPECTRAmax PLUS has been designed to regulate the temperature of the microplate chamber from 4 C above ambient to 45 C Upon power up when the incubator is off the temperature in the SPECTRAmax PLUS micro plate chamber is ambient and isothermal Pressing the incubator TEMP On Off key will cause the SPECTRAmax to begin warming the microplate cuvette chamber The temperature set point defaults to 37 0 C at start up Accuracy of the temperature set point is only guaranteed if the set point is at least 4 C above ambient If the temperature set point is lower than the ambient temperature the chamber temperature will remain at ambi ent Temperature regulation is controlled by heaters only and therefore cannot cool the tempe
46. sers Manual SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 15 nam Chapter 1 E Specifications Thermal specifications for microplates used in the SPECTRAmax PLUS apply to flat bottom microplates with isolated wells All other microplate specifications apply to standard 96 well polystyrene flat bottom microplates Performance spec ifications for cuvette readings apply only to aqueous solutions having solute molal concentrations less than 0 4 M When pathlength compensation is applied to microplate absorbance measurements agreement with cuvette absorbance mea surements for the same solution requires that the solution volume in the micro plate well be between 100 uL and 300 pL NONI Technical specifications are subject to change without notice Wavelength range Wavelength selection Wavelength bandwidth Wavelength accuracy Wavelength repeatability Photometric range Photometric resolution Photometric accuracy linearity microplate 0 2 0 OD Photometric accuracy linearity cuvette 0 2 0 OD Photometric precision repeatability 0 2 0 OD Stray light Photometric stabilization Photometric drift Calibration Optical alignment Light source Average lamp lifetime Illumination Photodetectors Photometric Performance 190 1000 nm Monochromator tunable in 1 nm increments lt 2 0 nm FWHM full width half maximum 1 0 nm across wavelength range 0 2 nm across all optical channels
47. stallation Warnings os ce ei Rh ao 2 3 Installation Cautions eo ROW VERE A a pus 2 3 a ee ea 0 eat 2 3 Setting Up for Stand Alone Use 2 4 Chapter 3 Stand Alone Operation 3 1 Prepare fora Reading tn af autom toe tp Mal 3 3 Turn the Instrument and Printer On 3 3 Set the Temperature osos da p 3 3 Select the Wavelength 3 3 Read the E CELE AWA 3 4 Operation Overview ia di 58 eaaa 3 4 SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual v Figures Table vi Chapter 4 Maintenance and Troubleshooting 4 1 Tecate eee ty 4 3 Warnings and Cautions 4 3 Generali a az Abe net dea ate tds 4 4 Cleaning 2 sa diea nb es an cdit eia 4 4 Changing the Fan Filter 4 4 Chene no dhe BUSES sao ee DA Eb cb ED dE ee 4 5 Error Codes and Probable Causes 4 7 Error Messages iii cue at ES M RT uns 4 7 Opening the Drawer Manually 4 11 Appendix A Printers and Cables A 1 Compatible PrintelSos ue ded te ata t
48. standards The PathCheck feature of the SPECTRAmax PLUS automatically determines the pathlength of aqueous samples in the microplate and normalizes the absorbance in each well to a pathlength of 1 cm This novel approach to Microplate Wells correcting the microwell absorbance values is accurate to within 2 5 of the values obtained directly in a 1 cm cuvette Reference measurements made by reading the cuvette or using factory stored val ues derived from deionized water can be used to normalize the OD data for microplate wells This pathlength correction is accomplished only with the use of SOFTmax PRO software Stand alone control of the instrument is limited to fixed wavelength 1 4 SPECTRAmax PLUS Microplate Spectropbotometer Operators Manual Instrument Description Component Description reference measurements made with the cuvette SOFTmax PRO software from Molecular Devices provides full instrument control and statistical data analysis The main components of the SPECTRAmax PLUS are The control panel The microplate drawer The cuvette chamber The back panel connections and power switch Ze Back Panel Control Panel Cuvette Chamber Microplate Drawer Figure 1 2 Major areas of the SPECTRAmax PLUS Control Panel Figure 1 3 Control Panel The control panel consists of an LCD and nine pressure sensitive color coded membrane keys which can be used to perform the stand a
49. t the right front of the SPECTRAmax PLUS the cuvette chamber has a lid that lifts up allowing you to insert or remove a cuvette The chamber contains springs that automatically position the cuvette in the proper alignment for a reading The cuvette door must be closed in order to initiate a reading Cuvettes The SPECTRAmax PLUS can accommodate standard height 45 mm 1 cm cuvettes and 12 x 75 mm test tubes when used with the test tube cover Figure 1 6 Test Tube Cover Not all manufacturers cuvettes are the same with regard to design materials or configuration Temperature uniformity within the cuvette may vary depending on the type of cuvette used Be sure to handle cuvettes on the frosted sides only Place the cuvette into the chamber so that the reading clear sides face left and right see Figure 1 5 The SPECTRAmax PLUS will not accept the short 25 mm high micro cuvettes SPECTRAmax PLUS Microplate Spectrophotometer Operators Manual 1 9 Chapter 1 O The Back Panel Power Switch y 3 Fuse Box Cover Power Cord AR Receptacle Printer Centronics Figure 1 7 Components on the Back Panel of the SPECTRAmax PLUS The following components are located on the back panel of the SPECTRAmax PLUS system Power switch a rocker switch labeled I O for on and off respectively Power cord receptacle plug the power cord in here Fuse box cover cannot be opened while the power cord
50. tech nological developments This manual is subject to change without notice Although this manual has been prepared with every precaution to ensure accuracy Molecular Devices Corporation assumes no liability for any errors or omissions nor for any damages resulting from the application or use of this in formation Molecular Devices Corporation Instrument Warranty Molecular Devices Corporation warrants this product against defects in material or workmanship as follows 1 All parts of the SPECTRAmax PLUS Microplate Spectrophotometer are warranted for a period of one 1 year from the original date of delivery 2 All labor charges to repair the product for a period of one 1 year from the original date of delivery will be paid by Molecular Devices Corporation 3 This warranty covers the SPECTRAmax PL US instrument only and does not extend to any computer printer software reagents or disposables used with the instrument Labor and Parts To obtain warranty service during the applicable warranty period you must take the product or deliver the product properly packaged in the original shipping materials and carton to an authorized Molecular Devices Corporation ser vice facility You must call or write to the nearest Molecular Devices Corporation service facility to schedule warranty service You may call Molecular Devices Corporation at the telephone number or address below to locate the nearest service facility You must schedule w
51. ual 33 Chapter 3 Read tbe Cuvette Operation Overview 3 After setting the temperature if desired and choosing the wavelength for the reading insert the cuvette into the chamber making sure that the clear sides are to the left and right facing the instrument Do not touch the clear surfaces of the cuvette Make sure the cuvette is completely seated in the chamber and close the cuvette door If the cuvette contains a blank typically this solvent contains everything that the samples contain except for analyte press the key to acquire the reference reading from the cuvette The SPECTRAmax PLUS will automatically calibrate in less than two seconds send a header for the reading to the printer close the microplate drawer if it was open and read the cuvette according to the selected instrument settings If the cuvette contains a sample touch the READ CUVETTE key to acquire the sample reading from the cuvette When reading is complete remove the cuvette The following steps provide a quick reminder of the basic operating procedures required to read a cuvette using the SPECTRAmax PLUS Turn on the power switch of the SPECTRAmax PLUS located on the back panel If you wish to regulate the temperature inside the microplate reading chamber touch the TEMP On Off key to bring the chamber to the default temperature of 37 0 C The microplate drawer will close and the indicator on the right of the LCD will f
52. ubleshooting information including error codes that may be seen followed by their most likely causes and remedies Molecular Devices Corporation is a leading worldwide manufacturer and distrib utor of analytical instrumentation We are committed to the quality of our prod ucts and to fully supporting our customers with the highest level of technical service In order to fully benefit from our technical services please complete the registration card and return it to the address printed on the card If you have any problems using the SPECTRAmax PLUS Microplate Spectro photometer that are not covered in this chapter in the U S contact our Techni cal Services group at 1 800 635 5577 elsewhere contact your local representative It is your responsibility to decontaminate the instrument as well as any accessories before requesting service by Molecular Devices representatives and before returning the instrument or any components to Molecular Devices Corporation AMINE All maintenance procedures described in this manual can be safely performed by qualified personnel Maintenance not covered in this manual should be performed by a Molecular Devices representative A EANNA Removal of protective covers that are marked with the High Voltage warning symbol shown below can result in a safety hazard A EZAR Always turn the instrument off and remove the power cord and any computer printer cables from the back prior to any
53. x PRO e Express data as Absorbance or Transmittance Single wavelength reading of microplate and or cuvette Multiple wavelength up to six reading of microplate and or cuvette Kinetic and kinetic graphics of microplate and or cuvette Spectral scan 190 1000 nm of microplate and or cuvette Measurement Time calibration off Microplate 96 wells in 9 seconds single wavelength 96 wells in 19 seconds dual wavelength 425 amp 650 nm 384 wells in 29 seconds single wavelength 384 wells in 59 seconds dual wavelength 425 amp 650 nm 96 wells in 5 seconds single wavelength 96 wells in 12 seconds dual wavelength 425 amp 650 nm 384 wells in 16 seconds single wavelength 384 wells in 34 seconds dual wavelength 425 amp 650 nm 96 wells 9 second minimum interval between readings single wavelength column 2 second minimum interval between readings single wavelength 384 wells 29 second minimum interval between readings single wavelength Cuvette second single wavelength 2 second minimum interval between readings single wavelength Measurement conditions endpoint column priority for dual wavelength measurements calibrate off Cuvette Normal scan Cuvette Speed scan Microplate Normal scan Microplate Speed scan Scan Speed 45 K nm min K wavelength interval 130 K nm min 33 K nm min 8 well strip 21 K nm min 16 well strip 135 K nm min
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