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1. D Ir 10 Reconnect the power cord to the instrument and to the wall oudet and reconnect other cables previously disconnected 28 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 6 1 Error Codes and Resolutions 6 Troubleshooting This chapter lists error codes that may occur while using the instrument followed by their most likely causes and remedies Maintenance procedures are described in Chapter 5 For problems with the SpectraMax or VersaMax that are not listed here in the U S contact Molecular Devices Technical Services group at 1 800 635 5577 elsewhere call your local representative N BIOHAZARD 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 6 1 Error Codes and Resolutions The LCD displays Fatal Error codes when a situation arises that requires attention Any reading in progress will stop Warning messages do not stop a reading 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 generally self explanatory For a
2. Operating temperature 5 C to 40 C Operating altitude lt 2000 m Installation category II Pollution degree 2 Operating humidity lt 80 Storage temperature 20 C to 65 C SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 47 Part 7 Specifications PHYSICAL Size hx wx d 8 6 220 mm x 22 8 580 mm x 15 380 mm Weight 30 Ib 13 6 kg Power consumption lt 250 W Line voltage and frequency 90 250 VAC autoranging 50 60 Hz 7 4 VersaMax Specifications Thermal specifications for microplates used in the VersaMax apply to flat bottom microplates with isolated wells All other microplate specifications apply to standard 96 well polystyrene flat bottom microplates Technical specifications are subject to change without notice PHOTOMETRIC PERFORMANCE Wavelength range 340 850 nm Wavelength selection Monochromator tunable in 1 nm increments Wavelength bandwidth lt 2 0 nm full width half maximum Wavelength accuracy 1 0 nm across wavelength range Wavelength repeatability 0 2 nm across all optical channels Photometric range 0 to 4 000 OD Photometric resolution 0 001 OD a S o o o 2 7 Photometric accuracy linearity 340 850 nm lt 1 0 and 0 006 OD microplate 0 2 0 OD Photometric precision repeata
3. gt o eo WARNING Never perform any operation on the instrument in an environment where liquids or potentially damaging gases are present WARNING Risk of electrical shock Refer servicing to qualified personnel A CAUTION Use of organic solvents such as dichloromethane may cause harm to the optics in the instrument Extreme caution is advised when using organic solvents Always use a plate lid and avoid placing a plate containing these materials in the read ing chamber for prolonged periods of time Damage caused by the use of incompati ble or aggressive solvents is NOT covered by the instrument warranty SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 23 o rr D i Part 5 Maintenance A CAUTION Never touch any of the optic mirrors filters or cables or their housing or manifold The optics are extremely delicate and critical to use of the instrument A CAUTION Do not touch or loosen any screws or parts other than those specifi cally designated in the instructions Doing so could cause misalignment and possibly void the warranty 5 2 Moving the VersaMax or SpectraMax 5 3 General If you need to relocate a VersaMax or SpectraMax follow these steps WARNING SpectraMax and VersaMax instruments weigh approximately 35 pounds 16 kilograms To avoid injury it is recommended that two people lift the instrument together using proper l
4. pa jas i ium N i E amp A N Max Pro manual 2 4 3 Pathcheck and Interfering Substances Any material that absorbs in the 900 nm to 1000 nm spectral region could interfere with PathCheck measurements Fortunately there are few materials that do interfere at the concentrations typically used Turbidity is the most common interference if you can detect any turbidity in your sample you should not use PathCheck Turbidity elevates the 900 nm measurement more than the 1000 nm measurement and causes an erroneously low estimate of pathlength Using a Cuvette Reference does not reliably correct for turbidity Samples that are highly colored in the upper visible spectrum may have absorbance extending into the near infrared NIR and can interfere with PathCheck Examples include Lowry assays molybdate based assays and samples containing hemoglobins or porphyrins In general if the sample is distinctly red or purple you should check for interference before using PathCheck To determine possible color interference do the following e Measure the optical density at 900 nm and 1000 nm both measured with air reference e Subtract the 900 nm value from the 1000 nm value 12 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 2 4 PathCheck not VersaMax e Do the same for pure water If the delta OD for the sample differs significantly from the delta OD for water th
5. 0112 0126 Rev A 3 i 7 a an a Part 1 Description in the front left corner of the drawer holds the plate securely in place when the drawer is closed The drawer remains in the reading chamber during read cycles Figure 1 2 The microplate drawer Microplate drawer operation varies depending on the incubator setting e Ifthe incubator is off the drawer remains open Ifthe incubator is on the drawer closes after approximately 10 seconds to assist in maintaining temperature control within the microplate chamber Do not obstruct the movement of the drawer If you must retrieve a plate after an error condition or power outage and the drawer does not open it is possible to open it manually see Section 6 Troubleshooting 1 2 3 Mlicroplates SpectraMax 190 and VersaMax readers can accommodate standard 96 well and strip well microplates SpectraMax Plus 84 and 340PC 4 readers can accommodate standard 96 well 384 well and strip well microplates When reading at wavelengths 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 4 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 1 2 Components 1 2 4
6. i oO 7 oO EI Pressing the up or down arrow key scrolls up or down through the available wave lengths starting at the previous setting e Pressing the up A or down W arrow once increments or decrements the dis played wavelength by 1 nm Pressing and holding either arrow increments or decrements the displayed wave length by 10 nm until it is released 96 384 SpectraMax 340PC only Selects either 96 well or 384 well plate for a stand alone read REF SpectraMax Plus 9 only 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 instrument uses the Ig values stored in the NVRAM non volatile memory of the instrument READ CUVETTE SpectraMax Plus 9 only Initiates the sample reading of the cuvette READ SpectraMax 340PC 84 and SpectraMax 190 only Initiates reading of the microplate in stand alone mode Data is sent to the printer port T A SpectraMax Plus 9 only A toggle switch used to display cuvette data as percent transmission or absorbance DRAWER The DRAWER key opens and closes the microplate drawer 1 2 2 The Microplate Drawer The microplate drawer is located on the right side of the SpectraMax and VersaMax readers slides in and out of the microplate chamber A small plastic pusher located SpectraMax and VersaMax Plate Readers Operator s Manual
7. 300 399 HARDWARE ERRORS thermistor faulty Unable to read a reasonable thermistor value Thermistor faulty or disconnected Main board problem or ambient tempera ture out of range safe temperature limit exceeded A temperature of over 50 C detected on one or more of the 4 thermistors Temper ature will be shut off and remain off until a successful completion of power up reset low 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 current Too much stray light detected on power up faulty or disconnected 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 light Minimum signal reference ratio not met during air calibration 32 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 6 1 Error Codes and Resolutions ERRORCODE ERROR MESSAGE NOTES 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 front panel not responding LCD front panel bad or disconnected gain calibration failed Power up calibration and check of signal path gain is out of tolerance Could be due to bad or disconnected
8. N e Wavelength control not VersaMax Fixed point microplate readings SpectraMax 190 and SpectraMax 340PC S e Fixed point cuvette readings SpectraMax Plus 84 2 5 1 Temperature Regulation SpectraMax and VersaMax instruments have been designed to regulate the tempera ture of the microplate chamber from 4 C above ambient to 45 C Upon power up when the incubator is off the temperature in the microplate chamber is ambient and isothermal Turning on the incubator by pressing the TEMP on off key causes the instrument to begin warming the microplate chamber The temperature set point defaults to 37 0 C at start up Accuracy of the temperature set point is guaranteed only 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 remains at ambient Temperature regulation is controlled by heaters only and therefore cannot cool the temperature to a setting lower than ambi ent Additionally the highest setting 45 C can be achieved only if the ambient tem perature is greater than 20 C Typically the microplate chamber reaches 37 0 C in less than 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 Should you turn the incubator back on after a momentary shutdown allow about ten minutes for the
9. The following components are located on the back panel of the SpectraMax and VersaMax readers 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 is plugged in When opened it provides access to the fuse box containing two fuses that are required for operation Computer port double shielded 8 pin RS 232 serial for use with an external computer plug one end of an 8 pin DIN serial cable into this port the other end attaches to the serial modem port of the computer Printer port double shielded 25 pin parallel for use in stand alone operation Plug the 25 pin end of the cable into this port the other end attaches to the port on the printer SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 1 2 Components Label provides information about the reader such as line voltage rating caution ary information serial number etc Record the serial number shown on this label for use when contacting Molecular Devices Technical Support m i oO 7 oO EI SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 7 Part 1 Description i 7 a 7 a 8 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 2 1 Absorbance 2 1 2 2 2 3 2 4 Principles of
10. k D E S z Endpoint Read In an endpoint read a reading of each microplate well is taken at a single or multiple wavelengths Depending on the read type selected values can be reported as optical density or 96 ransmittance Kinetic Read In a kinetic read the data are collected over time with multiple readings taken at regu lar intervals To achieve the shortest possible interval for kinetic readings choose wavelengths in ascending order Kinetic analysis can be performed for up to 99 hours The kinetic read interval depends upon the instrument setup parameters chosen in SoftMax Pro Kinetic analysis has many advantages when determining the relative activity of an enzyme in different types of microplate assays including ELISAs and the purifica tion and characterization of enzymes and enzyme conjugates Kinetic analysis is capable of providing improved dynamic range precision and sensitivity relative to endpoint analysis SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 15 Part 2 Principles of Operation Spectrum Read not VersaMax Spectral analysis measures optical density or Transmittance across a spectrum of wavelengths from 340 nm to 850 nm in the SpectraMax 340PC 84 190 nm to 850 nm in the SpectraMax 190 and 190 nm to 1000 nm in the SpectraMax Plus 94 All spectrum readings are made using the scanning monochromators of the instrument 2 5 3 Automix The Aut
11. 250 W Line voltage and frequency 90 250 VAC autoranging 50 60 Hz 7 2 SpectraMax 340PC Specifications Thermal specifications for microplates used in the SpectraMax 340P cs apply to flat bottom microplates with isolated wells All other microplate specifications apply to standard 96 well polystyrene flat bottom microplates Technical specifications are subject to change without notice PHOTOMETRIC PERFORMANCE Wavelength range 340 850 nm Wavelength selection Monochromator tunable in 1 nm increments Wavelength bandwidth lt 2 0 nm full width half maximum Wavelength accuracy 1 0 nm across wavelength range Wavelength repeatability 0 2 nm across all optical channels Photometric range 0 to 4 000 OD Photometric resolution 0 001 OD Photometric accuracy linearity microplate 0 2 0 OD 340 850 nm 1 0 and 0 006 OD 40 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 7 2 SpectraMax 340PC Specifications Photometric precision repeatability 0 2 0 OD 340 850 nm 1 0 and 0 003 OD Stray light 0 0596 at 340 nm Photometric stabilization Instantaneous Photometric drift None continuous referencing of monochromatic input Calibration Automatic before first kinetic read and before every endpoint reading Optical alignment None required L
12. 7 Part 7 Specifications TEMPERATURE REGULATION Reading chamber Isothermal when temperature regulation is not enabled Range 4 C above ambient to 45 C when tempera ture regulation enabled The ambient tem perature must be gt 20 C to achieve temperature regulation at 45 C Resolution 0 1 C 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 continu ously 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 between kinetic readings Plate mixing duration Selectable 1 to 999 seconds three second default 38 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 7 1 SpectraMax Plus Specifications COMPATIBILITY Microplates Standard and half area 96 well flat bot tomed microplates 0 3 mL 384 well flat bottomed microplates Polystyrene plates for wavelengths abov
13. The Cuvette Chamber SpectraMax Plus only m i oO 7 oO EI Figure 1 3 The cuvette chamber Located at the right front of the SpectraMax Plus 84 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 before initiating a reading Cuvettes The SpectraMax Plus 84 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 4 Not all manufacturers cuvettes are the same with regard to design materials or con figuration Temperature uniformity within the cuvette may vary depending on the 8 y vary dep 8 type of cuvette used Be sure to handle cuvettes on the frosted sides only Place the cuvette into the cham ber so that the reading clear sides face left and right The SpectraMax Plus 8 does not accept the short 25 mm high microcuvettes SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA i 7 a 7 a Part 1 Description 6 Figure 1 4 Test tube cover 1 2 5 The Back Panel Fuse Box Cover Power Switch Power Cord Receptacle e Figure 1 5 Schematic of the back panel of a reader Computer Printer Port Port
14. mea Instrument could not perform command surement in progress because a measurement was in progress 107 no data to transfer Inputting transfer when there s no data in the buffer 108 data buffer full Too 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 cuvette door Cuvette door open while doing a read open 111 invalid read settings 200 299 FIRMIWARE 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 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 31 o gt n im Part 6 Troubleshooting D i lt N 2 im o serial port parity error Parity bit error detected with incoming serial data serial port overrun error Caused by host computer sending too much data and ignoring the flow control signal serial port framing error cmd generated too much output Firmware error fatal trap Instrument error Instrument locks up RTOS error Firmware error stack overflow Firmware error unknown interrupt Firmware error
15. 2 4 PathCheck not VersaMax 00 00 eee ete eee 9 2 4 1 Water Constant or Cuvette Reference Option SpectraMax Plus s Only bens Ee a tute papae 11 2 4 2 Background Constant Subtraction and Blanking Considerations 12 2 4 3 Pathcheck and Interfering Substances 0 0 e eee eee 12 2 4 4 Making Absorbance Measurements Normalized to a 1 cm Pathlength 13 2 5 Functional Description 2 00 eee 14 2 5 1 Temperature Regulation ii gns aske ions ae Peewee eae 14 2 5 2 Read Iypesz i eo bere td beatae eco EP nds to Er tee a 15 BSD AUTORE eee ab Edd e p E REM RR i 16 2 5 4 Computer Control ag DA ek Top Ea PERPE REFER eee nid gay 16 3 Installati n evi Eier ER ea aciei d ui abok dol o MODA beaten 17 3 1 Unpacking i d x E Ses ee G eed Pw ede ERIG bx RR E ERU PASA 17 3 2 Setting Up the Instrument 00 0 cee eee 17 4 MOSH eh OW ear ce ae canine co DT TIMES 19 4 1 Quick Overview ioci tl gg tbed ei bem abet E xad Goede bie bane 19 4 2 Preparing for a Reading 0c cece eee eens 19 4 2 1 Turn the Instrument and Computer On 0 0 000 eee eee ee 19 4 2 2 Set the Temperature Optional 0 0 eee eee cee 20 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA i Contents ii 4 3 Read the Microplate 00 0 4 4 Standalone Endpoint Cuvette Reading SpectraMax Plus 84 Only eeu Baten entere pauco Seated wn
16. 84 Specifications Thermal specifications for microplates used in the SpectraMax Plus s bottom microplates with isolated wells apply to flat All other microplate specifications apply to standard 96 well polystyrene flat bottom microplates Performance specifications for cuvette readings apply only to aqueous solutions hav ing solute molal concentrations less than 0 4 M When pathlength compensation is applied to microplate absorbance measurements agreement with cuvette absorbance measurements for the same solution requires that the solution volume in the microplate well is between 100 uL and 300 uL Technical specifications are subject to change without notice PHOTOMETRIC PERFORMANCE Wavelength range 190 1000 nm Wavelength selection Monochromator tunable in 1 nm increments Wavelength bandwidth lt 2 0 nm full width half maximum Wavelength accuracy 1 0 nm across wavelength range Wavelength repeatability 0 2 nm Photometric range 0 to 4 000 OD Photometric resolution 0 001 OD Photometric accuracy linearity 190 1000 nm lt 1 0 and 0 006 OD microplate 0 2 0 OD zx 4 e Lr O D 7 Photometric accuracy linearity 190 1000 nm lt 1 0 and 0 005 OD cuvette 0 2 0 OD Photometric precision repeatability 190 1000 nm lt 1 0 and 0 003 OD 0 2 0 OD Stray light lt 0 05 at 230 nm Photometri
17. As shown in the figures below press to the left 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 Power Power Receptacle Fuse Box Switch Figure 5 1 Power receptacle fuse box and power switch 5 Continue gently pulling the fuse box forward until it is free of the instrument 26 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 5 6 Changing the Fuses Figure 5 2 Removing the fuse box 6 When removed the fuse assembly will appear as shown in Figure 5 3 The holder inside contains two fuses m m gt o eo Figure 5 3 The fuse box and holder with fuses removed from instrument SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 27 Part 5 Maintenance Figure 5 4 The fuse holder with fuses removed from the fuse box 7 It is possible that only one of the fuses may have blown However Molecular Devices recommends that you replace both fuses to ensure continued proper operation Pull both fuses out of the holder and discard them 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 left side toward the power receptacle Press the fuse box into place making sure the cover snaps closed o rr
18. Operation Absorbance Absorbance is the amount of light absorbed by a solution To measure absorbance accurately it is necessary to eliminate light scatter In the absence of turbidity absor bance optical density A log I D where Jp is incident light and Zis transmitted light In this manual we use the terms absorbance and optical density interchangeably Optical Density Optical density is 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 sam ple plus light scatter from turbidity Transmittance Transmittance is the ratio of transmitted light to the incident light T Iy I T 100T where Jp is incident light and J is transmitted light PathCheck not VersaMax The Beer Lambert law states that absorbance is proportional to the distance that light travels through the sample A bc where A is the absorbance is the molar absorbtivity of the sample 6 is the path length and c is the concentration of the sample In short the longer the pathlength 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 perform extinction based assays and also makes it confusing to compare results between microplate readers and spectrophotometers The standard pathlength of a cuvette i
19. control algorithm to fully stabilize the microplate chamber temperature 14 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 2 5 Functional Description Temperature regulation and control of the microplate chamber is achieved through electric heaters a fan efficient insulation and temperature sensors The heaters are located in the microplate chamber which is insulated to maintain the temperature set point The sensors are mounted inside the chamber and measure the air temperature The temperature feedback closed loop control algorithms measure the chamber air temperature compare it to the temperature set point and use the difference to calcu late the regulation of the heating cycles This technique 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 temperature uniformity within the microplate depends on its design and composition Note the temperature display on the SpectraMax Plus 4 control panel shows the temper ature of the cuvette chamber The microplate chamber temperature can be seen only using SoftMax Pro 2 5 2 Read Types SpectraMax and VersaMax readers can perform up to three types of read depending on the instrument model endpoint kinetic and spectrum Instrument setup parame ters for each read type are discussed in the SoftMax Pro User s Manual p mz 9 D 7
20. initiation of temperature regulation Temperature regulation 4 sensors Drift 0 2 C regulated Temperature regulation diagnostics Temperature regulation system is continu ously monitored and updated Evaporation Plate lid required to minimize evaporative cooling SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 7 3 SpectraMax 190 Specifications 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 between kinetic readings Plate mixing duration Selectable 1 to 999 seconds three second default COMPATIBILITY Microplates Standard and half area 96 well flat bot tomed microplates 0 3 mL GENERAL INSTRUMENT Display 2 x 20 character backlit LCD Operating panel 8 key membrane keypad Self diagnosis Continuous on board diagnostics Spill control Drawer mechanism and reading chamber assembly protected from accidental spillage by drainage ports Computer interface 8 pin DIN RS 232 serial double shielding required Printer interface Parallel 25 pin to Centronics double shielding required Microplates supported All 96 well and strip well microplates including lids ENVIRONMENTAL FOR INDOOR USE ONLY zx 4 e Lr O 9 7
21. unreasonable data error 503 NVRAM Plate air cal data Plate air calibration data is unreasonable error 504 NVRAM Carriage offset Carriage offset data is unreasonable error 505 NVRAM Stage offset error Stage offset data is unreasonable 506 NVRAM Battery Time to replace the NVRAM battery U3 For all other error messages codes not listed here please contact your local Molecu lar Devices representative for assistance 6 2 Opening the Drawer Manually fan error occurs while the drawer is closed and you need to remove a microplate press the DRAWER key Ifthe drawer does not open turn power to the instrument off and then on again Ifthe drawer remains closed turn the incubator off if it was on by pressing the TEMP on off key Ifthe drawer still remains closed try using a blunt flat object such as a spatula to open the door 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 D i i N 2 EI If you are still unable to open the drawer contact your local Molecular Devices representative 34 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 7 1 SpectraMax Plus 9 Specifications 7 Specifications 7 1 SpectraMax Plus
22. 36 Power Cord EC2 UK Indonesia Singapore Malaysia 4400 0037 Power Cord AP1 Australia Hong Kong China 4400 0038 Fuse 4 amp 5 x 20 mm Time Delay 4601 0014 SpectraMax Mouse Pad 9000 0133 Adapter USB Serial High Speed KeySpan adapter instrument to USB 9000 0938 only instrument Test Tube Cover 2300 0277 8 3 Cuvettes in SpectraMax Plus 94 The guidelines for cuvette use in the SpectraMax Plus 84 are the same as those that apply to any high quality spectrophotometer The user must ensure that the meniscus is comfortably above the light beam in standard cuvettes and that the sample chamber in a microcuvette is aligned properly with the beam The light beam is 0 625 in 15 87 mm above the cuvette bottom Below are some cuvettes that have been tested All have an optical pathlength of 1 cm 10 mm and standard external dimensions 12 5 cm x12 5 cm Their fill volumes differ only because of their different internal width and chamber height dimensions P o 5 o 2 2 lt co 54 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 8 3 Cuvettes in SpectraMax Plus 384 Standard and Semi micro Cuvettes Several brands available Internal Width Minimum Volume Maximum Volume 10 mm 1 8 mL 4 0 mL 4mm 0 75 mL 1 4 mL 2mm 0 40 mL 0 7 mL Ultra micro Cuvettes Hellma When ordering specify the Z dimension to be 15 mm Hell
23. 6 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 Microplate read time endpoint speed read Measurement conditions endpoint column priority for dual wavelength measurements calibrate off 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 Microplate kinetic read intervals 96 wells 9 second minimum interval between readings single wavelength 1column 2 second minimum interval between readings single wavelength 384 wells 29 second minimum interval between readings single wavelength Cuvette read time endpoint 1 second single wavelength Cuvette kinetic read intervals 2 second minimum interval between readings single wavelength SCAN SPEED Cuvette normal scan 45 K nm min K wavelength interval Cuvette speed scan 130 K nm min Microplate normal scan 33 K nm min 8 well strip 21 K nm min 16 well strip Microplate speed scan 135 K nm min 8 well strip 77 K nm min 16 well strip SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 37 zx c e Lr O D 7 a S o o o 2
24. Cee OO 4 5 Standalone Endpoint Microplate Reading SpectraMax 190 and SpectraMax 340PC 9 Only 0 nens une naear Maintenance cenis tete epERER ERR debi eve Lx RM FREE Rae 5 1 Technical Support 4258 lRkgu cad RR eee eee eed EH EEG Lea E 5 2 Moving the VersaMax or SpectraMax 002 0c eee ete ee 5 3 General Maintenance 00 cece nnn 5 4 Cleaning eor bRE ERI ID RR AEn hac deles ieee amet dues es Oed 5 5 Cleaning the Fan Filter 00 0 cece eee ete eh 5 6 Changing the Fuses 2 c eR EIE WR ae ots Rede ee ee ERE DRE Troubleshootlrig 2 oe ROC er ir ende E RERO ER ER Ie En acu api s 6 1 Error Codes and Resolutions 00 cece eee eee eee elles 6 1 1 Error Code Classifications seses RR enn enn eens 6 2 Opening the Drawer Manually 0000 eee Specifications 24 wc dccesn ke ore ies Semen de dbs dee 7 1 SpectraMax Plus 4 Specifications 7 2 SpectraMax 340PC 9 Specifications 00 ccc cece eee cence renere 7 3 SpectraMax 190 Specifications 0000 ccc eee 7 4 VersaMax Specifications 00 cece eee Appendices 15 c ETT 8 1 Cables 2 4 23 25233ie2c ie cia a RPG pb GbMEESRISR MS BRE ueDede ipe 8 2 ACCcessOrleS oni ors E ERI quedo virtud sut a Hae a Rer d See Du PER 8 3 Cuvettes in SpectraMax Plus 84 FASE tate sere eb ee dee eee Dhaene SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 1 1 Intr
25. SpectraMax Plus 94 SpectraMax 190 SpectraMax 340PC 4 VersaMax Microplate Spectrophotometer Operator s Manual Molecular Devices Molecular Devices Corporation 1311 Orleans Drive Sunnyvale California 94089 Part 0112 0126 Rev A SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A Molecular Devices Corporation SpectraMax Plus3 SpectraMax 190 SpectraMax 340PC384 and VersaMax Operator s Manual Copyright Copyright 2005 Molecular Devices Corporation All rights reserved No part of this publication may be reproduced 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 4 SpectraMax 190 SpectraMax 340PC284 VersaMax and methods are protected by the following patents U S Patents 4 968 148 5 112 134 6 151 111 and 6 188 476 Canadian Patent 1 305 334 Other U S and International pat ents pending Trademarks SpectraPlate PathCheck and Automix are trademarks and SpectraMax and SoftMax are registered trademarks of Molecular Devices Corporation Apple and Macintosh are registered trademarks of Apple Computer Incorporated Windows is a registered tra
26. ance between 900 nm and 1000 nm at concentrations normally used PathCheck can also be used with samples containing small amounts of organics or high buffer concentrations by using the Cuvette Reference below 2 4 1 Water Constant or Cuvette Reference Option SpectraMax Plus384 Only The PathCheck measurement is based on the absorbance of water in the near infrared region between 900 nm and 1000 nm If the sample is completely aqueous has no turbidity and has a low salt concentration less than 0 5 M the Water Constant is adequate The Water Constant is determined during manufacture and is stored in the instrument If the sample contains an organic solvent such as ethanol or methanol we recom mend using the cuvette reference It is important that the solvent does not absorb in the 900 nm to 1000 nm range to determine whether or not a given solvent would interfere see the discussion of interfering substances below When a noninterfering solvent is added to the aqueous sample the water absorbance decreases proportionally to the percentage of organic solvent present For example 5 ethanol decreases the water absorbance by 5 and results in a 5 underestimation of the pathlength You can avoid the error by putting the same water solvent mixture in a cuvette and using the Cuvette Reference To use the Cuvette Reference place into the cuvette port a standard 1 cm cuvette containing the aqueous solvent mixture that is used for the sampl
27. bility 340 850 nm 1 0 and 0 003 OD 0 2 0 OD Stray light lt 0 0596 at 340 nm Photometric stabilization Instantaneous 48 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 7 4 VersaMax Specifications Photometric drift None continuous referencing of monochromatic input Calibration Automatic before first kinetic read and before every endpoint reading Optical alignment None required Light source Xenon flash lamp 5 Watts Average lamp lifetime 1 billion flashes Illumination Top down Photodetectors Silicon photodiode PHOTOMETRIC ANALYSIS MODES Using SoftMax Pro Express data as Absorbance or 96 Transmittance Single wavelength reading of microplate Dual wavelength reading of microplate Kinetic and kinetic graphics of microplate MEASUREMENT TIME CALIBRATION OFF Read time endpoint standard read 96 wells in 9 seconds single wavelength 96 wells in 19 seconds dual wavelength 425 amp 650 nm Kinetic read intervals 96 wells 9 second minimum interval between readings single wavelength 1column 2 second minimum interval between readings single wavelength zx 4 e Lr O 9 7 TEMPERATURE REGULATION Reading chamber Isothermal when temperature regulation is not enabled Range 4 C above ambient to 45 C when tempera tur
28. ble the incubator Press the incubator TEMP on off key The LCD display indicates that temperature control is on and shows the set point and current temperature of the microplate chamber To change the temperature set point Press the up or down arrow keys until the desired temperature set point is shown in the display L eo 2 i The microplate chamber temperature is maintained at the set point until you disable temperature control by touching the incubator key again When the incubator is off the temperature within the microplate chamber gradually returns to ambient Should you turn the incubator back on after a momentary shutdown allow about ten minutes for the control algorithm to fully stabilize the microplate chamber temperature 4 3 Read the Microplate A BIOHAZARD The underside of the microplate must be dry prior to placing it in the drawer If the microplate has fluid on the underside dry it using a paper towel or equivalent before placing it in the drawer Insert the filled microplate into the drawer matching well Al with the upper left hand position of the drawer Make sure the microplate is flat against the drawer bottom You must have SoftMax Pro software running on a computer connected to the instrument Press the READ button in SoftMax Pro to start the plate read When reading is complete the drawer of the instrument opens allowing you to remove the m
29. c stabilization Instantaneous SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 35 a S o o o 2 7 Part 7 Specifications Photometric drift None continuous referencing of monochromatic input Calibration Automatic before first kinetic read and before every endpoint reading Optical alignment None required Light source Xenon flash lamp 5 Watts Average lamp lifetime 1 billion flashes Illumination Photodetectors PHOTOMETRIC ANALYSIS MODES Top down microplates horizontal cuvettes Silicon photodiode Standalone Single wavelength Absorbance or Transmittance reading of the cuvette or test tube Using SoftMax Pro Express data as Absorbance or Transmittance Single wavelength reading of micro plate and or cuvette Multiple wavelength up to six reading of microplate or cuvette Kinetic and kinetic graphics of micro plate and or cuvette Spectral scan 190 1000 nm of micro plate and or cuvette 36 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 7 1 SpectraMax Plus Specifications MEASUREMENT TIME CALIBRATION OFF Microplate read time endpoint standard read Measurement conditions endpoint column priority for dual wavelength measurements calibrate off 96 wells in 9 seconds single wavelength 9
30. cence and the ability of SoftMax Pro software to print on PC compatible and Macin tosh compatible printers Molecular Devices does not recommend relying on printing in standalone mode Keep the cuvette door closed when reading a cuvette See Appendix 8 3 Cuvettes in SpectraMax Plus384 for cuvette types that have been tested with the SpectraMax Plus ee 4 5 Standalone Endpoint Microplate Reading SpectraMax 190 and SpectraMax 340PC 9 Only When operating the SpectraMax 190 and SpectraMax 340PC 84 as stand alone Sys tems you can obtain a fixed wavelength reading of the microplate SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 21 Part 4 Operation e Choose the wavelength by pressing the up or down arrows beneath the display labeled A Set the temperature of the microplate chamber using the keys on the left side of the control panel To read the microplate e Press the key The resulting data is sent to the printer port of the instrument Note due to the continuous changes in printer technology leading to printer obsoles cence and the ability of SoftMax Pro software to print on PC compatible and Macin tosh compatible printers Molecular Devices does not recommend relying on printing in standalone mode o dem gt i 22 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 5 1 Technical Support 5 Maintenance 5 1 Tec
31. d have been placed in the packing carton Make sure all these items are present before proceeding 3 2 Setting Up the Instrument 1 Place the instrument on a level surface away from direct sunlight dust drafts vibration and moisture Turn the instrument around so that the back of the instrument is facing you Insert the serial cable into the RS 232 serial port receptacle on the back panel of the instrument A Keyspan USB adapter is necessary for a Macintosh computer or a Windows computer without a serial port see Appendix 8 1 Cables for more infor mation on adapter cables Attach the other end to your computer SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 17 e an 2 2 i z 2 T 4 en Part 3 Installation Insert the female end of the power cord into the power receptacle at the rear of the instrument 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 microplates in stand alone mode locate the printer port 25 pin parallel on the rear panel Connect one end of the cable to this port and connect the other end to the printer Load paper into the printer according to the manufacturer s instructions and connect the printer s power cord Turn the instrument around
32. demark of Microsoft Corporation 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 technological 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 information SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA Contents Contents 1 Description oic we EERERIRAT A bpeRGdbRPe heed deans dear eeosaaa 1 1 1 Introd ction 2 ede ieee eee SOEUR ACE cic CIERRE 1 1 2 Components ccuss o Ei IIXERRGR eh bee IER E eee eee bs be eas 1 LT Ehe Control Panel 25252225 dco Re E ER ERA dei aa ede a 2 1 2 2 The Microplate Drawer 2 eect eee 3 L GE Microplates 22 ii eT esp tide titan vent dda eis eee ed 4 1 2 4 The Cuvette Chamber SpectraMax Plus 8 only 5 1 2 5 The Back Panel in REP be Bad Rae Ae 6 2 Principles of Operation 0 0 0 ccc eee e cee eee ees 9 2 1 Absorbance se She a sole edie oho Buc Peed stesura lectu Rea ecto turus Eh pow Sed lena dee 9 2 2 Optical Density coco ue oe ee ee ee RR ea 9 2 3 Transmittance oce aue nux eR ode eRe el Sli we ee oe 9
33. e 340 nm UV transpar ent plates above 220 nm quartz plates above 190 nm Cuvettes Standard height 45 mm cells with 10 mm pathlength 12 5 mm x 12 5 mm outside with minimum inside width of 4 mm typical for 3 mL volume cells See Section 8 3 Cuvettes in SpectraMax Plus384 for more information Test tubes 12 x 75 mm test tubes can be used in the cuvette chamber with the test tube cover GENERAL INSTRUMENT Display Operating panel Self diagnosis Spill control 2 x 20 character backlit LCD 8 key membrane keypad Continuous on board diagnostics Drawer mechanism and reading chamber assembly protected from accidental spillage by drainage ports Computer interface 8 pin DIN RS 232 serial double shielding required Printer interface Parallel 25 pin to Centronics double shielding required Microplates supported All 96 well and strip well microplates including lids SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 39 zx 4 e Lr O rt 7 a S o o o 2 7 Part 7 Specifications ENVIRONMENTAL Operating temperature 15 C to 40 C Operating humidity 0 to 70 non condensing Storage temperature 20 C to 65 C PHYSICAL Size h x w x d 8 6 220 mm x 22 8 580 mm x 15 380 mm Weight 30 Ib 13 6 kg Power consumption lt
34. e regulation enabled The ambient tem perature must be gt 20 C to achieve temperature regulation at 45 C SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 49 a S o o o 2 7 Part 7 Specifications Resolution 0 1 C Accuracy 1 0 C for microplate 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 continu ously monitored and updated Evaporation Recommended microplate Plate lid required to minimize evaporative cooling Flat bottom microplates with isolated wells and lid AUTOMIX WITH SOFTMAX PRO Plate mixing modes Plate mixing duration Selectable off once prior to any reading and once prior to and between kinetic readings Selectable 1 to 999 seconds three second default COMPATIBILITY Microplates Standard and half area 96 well flat bot tomed microplates 0 3 mL GENERAL INSTRUMENT Display 2 x 20 character backlit LCD Operating panel 8 key membrane keypad Self diagnosis Continuous on board diagnostics Spill control Computer interface Drawer mechanism and reading chamber assembly prot
35. ected from accidental spillage by drainage ports 8 pin DIN RS 232 serial double shielding required 50 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 7 4 VersaMax Specifications Printer interface Parallel 25 pin to Centronics double shielding required Microplates supported All 96 well and strip well microplates including lids ENVIRONMENTAL FOR INDOOR USE ONLY Operating temperature 5 C to 40 C Operating altitude lt 2000 m Installation category II Pollution degree 2 Operating humidity 8096 Storage temperature 20 C to 65 C PHYSICAL Size h x w x d 8 6 220 mm x 22 8 580 mm x 15 380 mm Weight 30 Ib 13 6 kg Power consumption lt 250 W Line voltage and frequency 90 250 VAC autoranging 50 60 Hz SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 51 C e O O 9 7 Part 7 Specifications a S o o o 2 7 52 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 8 1 Cables 8 Appendices 8 1 Cables Molecular Devices recommends that you use high quality double shielded cables to connect your SpectraMax or VersaMax reader to the computer Choose cables that meet the following requirements Serial Interface Cable The serial interface cable used
36. ectra Max Plus 94 only or using factory stored values derived from deionized water Water Constant can be used to normalize the optical density data for microplate wells PathCheck pathlength correction is accomplished only when using the SoftMax Pro software The SpectraMax Plus and SpectraMax Plus offer both the Cuvette Reference and the Water Constant methods The SpectraMax 190 and SpectraMax 340P C 84 use the Water Constant only The actual pathlength Z of a solvent is found from the following equation Sample OD 1995 OD 900 k d cm 10 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 2 4 PathCheck not VersaMax When a Cuvette Reference is used for pathlength correction the value of k is obtained by taking optical density measurements on the fluid in the cuvette at two wavelengths 1000 and 900 nm k Cuvette OD i0007 OD ga When the Water Constant is used for pathlength correction the value of k is obtained from the instrument This constant is saved in the instrument in the factory and may differ slightly from instrument to instrument Once the pathlength d is found the following equation is used for the pathlength correction OD OD sample cm d cm pL 0 9 7 k te i 5 PathCheck is applicable to almost all biological pharmaceutical molecules in aque ous solution because they have little or no absorb
37. en it is advisable not to use the PathCheck feature Use of Cuvette Reference does not correct for the interference with the current calculation scheme in SoftMax Pro Cur rently Cuvette Reference involves a single automated read at 900 nm and 1000 nm and the automated calculations in SoftMax Pro do not compensate for color or sol vent interference However you could correct for such interference by taking two cuvette measurements and using a different set of calculations For further informa tion contact Molecular Devices Technical Support Organic solvents could interfere with PathCheck if they have absorbance in the region of the NIR water peak Solvents such ethanol and methanol do not absorb in the NIR region so they do not interfere except for causing a decrease in the water absorbance to the extent of their presence in the solution Their passive interference N me e 2 gt i 3 t F i can be avoided by using the Cuvette Reference If however the solvent absorbs between 900 and 1000 nm the interference would be similar to the interference of highly colored samples described above If you are considering adding an organic sol vent other than ethanol or methanol you are advised to run a spectral scan between 900 nm and 1000 nm to determine if the solvent would interfere with PathCheck 2 4 4 Making Absorbance Measurements Normalized to a 1 cm Pathlength SoftMax Pro automa
38. es above 220 nm quartz plates above 190 nm GENERAL INSTRUMENT Display 2 x 20 character backlit LCD Operating panel 8 key membrane keypad Self diagnosis Continuous on board diagnostics Spill control Drawer mechanism and reading chamber assembly protected from accidental zx 4 e Lr O D CO spillage by drainage ports Computer interface 8 pin DIN RS 232 serial double shielding required Printer interface Parallel 25 pin to Centronics double shielding required Microplates supported All 96 well and strip well microplates including lids SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 43 a S T o o o 2 7 Part 7 Specifications ENVIRONMENTAL FOR INDOOR USE ONLY Operating temperature 5 C to 40 C Operating altitude lt 2000 m Installation category II Pollution degree 2 Operating humidity lt 80 Storage temperature 20 C to 65 C PHYSICAL Size h x w x d 8 6 220 mm x 22 8 580 mm x 15 380 mm Weight 30 Ib 13 6 kg Power consumption lt 250 W Line voltage and frequency 90 250 VAC autoranging 50 60 Hz 7 3 SpectraMax 190 Specifications tom microplates with isolated wells microplates Thermal specifications for microplates used in the SpectraMax 190 apply to flat bot All o
39. es in the microplate The cuvette must be in place when you read the microplate When you click the Read SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 11 Part 2 Principles of Operation button in SoftMax Pro the instrument first makes the 900 nm and 1000 nm mea surements in the cuvette and then makes the designated measurements in the micro plate The cuvette values are stored temporarily and used in the PathCheck calculations for the microplate samples Use of Cuvette Reference with PathCheck is different from a reference reading of a cuvette in a CuvetteSet section by clicking the Ref button in the CuvetteSet section tool bar in SoftMax Pro The cuvette reference used for PathCheck calculations measurements at 900 nm and 1000 nm does not produce data that can be viewed in a CuvetteSet section and is used only with data in microplates not cuvettes 2 4 2 Background Constant Subtraction and Blanking Considerations Raw optical density measurements of microplate samples include both pathlength dependent components sample and solvent and a pathlength independent com ponent OD of microplate material The latter must be eliminated from the Path Check calculation in order to obtain valid PathCheck normalized results There are three ways to accomplish this plate blanks plate background constants and plate pre reads all of which are described in the PathCheck section of the Soft
40. hnical Support Molecular Devices Corporation is a leading worldwide manufacturer and distributor of analytical instrumentation We are committed to the quality of our products and to fully supporting our customers with the highest possible 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 your SpectraMax or VersaMax reader in the U S contact our Technical Services group at 1 800 635 5577 elsewhere contact your local representative N BIOHAZARD 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 WARNING All maintenance procedures described in this manual can be safel P y performed by qualified personnel Maintenance not covered in this manual should be performed only by a Molecular Devices representative WARNING Removal of protective covers that are marked with the High Voltage warning symbol shown below can result in a safety hazard AN WARNING Always turn the power switch off and disconnect the power cord from the main power source before performing any maintenance procedure that requires removal of any panel or cover or disassembly of any interior instrument component m m
41. icroplate If the incubator is on the drawer closes again after approx imately 10 seconds 20 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 4 4 Standalone Endpoint Cuvette Reading SpectraMax Plus 8 Only Ifyou return to the instrument and find the drawer closed after a reading has finished press the DRAWER key When the drawer opens you can remove the microplate For more information about plate reading please consult the SoftMax Pro User s Manual 4 4 Standalone Endpoint Cuvette Reading SpectraMax Plus 9 Only When operating the SpectraMax Plus fixed wavelength reading of the cuvette 384 45 a stand alone system you can obtain a Choose the wavelength by pressing the up or down arrows beneath the display labeled A Set the temperature of the cuvette microplate chamber using the keys on the left side of the control panel To read the cuvette m k t 5 Press the REF key for a reference reading Press 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 The data is sent to the printer port of the instrument A new cuvette can be loaded into the SpectraMax Plus 9 while the results from the first reading are being printed Note due to the continuous changes in printer technology leading to printer obsoles
42. ifting techniques Remove any microplate from the drawer and then close the drawer Turn off the power switch and unplug the power cord from the source and from the receptacle on the back of the instrument Depending on the distance that you are moving the instrument you may want to repackage the instrument in its original shipping carton Otherwise carry the instru ment or place it on a rolling cart to transport it Ensure that the new location meets the proper specifications as described in Section 3 2 Setting Up the Instrument Maintenance Keep the drawer closed when the instrument is not in use The drawer can be opened by pressing the DRAWER button Always close the drawer immediately prior to switching the instrument off 5 4 Cleaning N BIOHAZARD Wear gloves during any cleaning procedure that could involve con tact with either hazardous or biohazardous materials or fluids WARNING Never clean the inside of the instrument Periodically you should clean the outside surfaces of the instrument 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 24 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 5 5 Cleaning the Fan Filter Do not spray cleaner directly onto the instr
43. ight source Xenon flash lamp 5 Watts Average lamp lifetime 1 billion flashes Illumination Top down Photodetectors Silicon photodiode PHOTOMETRIC ANALYSIS MODES Standalone Single wavelength Absorbance read ing of the microplate Using SoftMax Pro Express data as Absorbance or Transmittance Single wavelength reading of microplate Multiple wavelength up to six reading of microplate Kinetic and kinetic graphics of microplate Spectral scan 340 850 nm of microplate SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 41 z 74 e Lr O D 7 a S o o o 2 7 Part 7 Specifications MEASUREMENT TIME CALIBRATION OFF Read time endpoint standard read 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 Kinetic read intervals 96 wells 9 second minimum interval between readings single wavelength 1column 2 second minimum interval between readings single wavelength e 384 wells 29 second minimum interval between readings single wavelength SCAN SPEED Microplate normal scan 33 K nm min 8 well strip 21 K nm min 16 well strip Microplate speed scan 135 K nm min 8 well stri
44. m the back of the instrument prior to any installation or relocation of the instrument WARNING Do not operate the instrument in an environment where potentially damaging liquids or gases are present A CAUTION Do not touch or loosen any screws or parts other than those specifi cally designated in the instructions Doing so might cause misalignment and voids the instrument warranty 3 1 Unpacking SpectraMax and VersaMax instruments are 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 ship ping 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 A WARNING SpectraMax and VersaMax instruments weigh approximately 30 pounds 13 6 kg and should be lifted with care It is recommended that two persons 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 instrument 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 The packing list that accom panies the instrument describes all components that shoul
45. ma Cat No Window Size Chamber Volume Fill Volume 105 201 OS 2 0 x 5 0 mm 100 uL 120 uL 105 202 0S4 2 0 x 2 5 mm 50 uL 70 uL 105 210 QS gt 0 8 mm diameter 5 ul 10 uL a You must put a riser 0 81 mm on cuvette bottom to match the cuvette window to the beam b You must put a riser 0 8 1 mm on cuvette bottom to match the cuvette window to the beam Gives good qualitative results i e spectral scans but quantitative results are impractical because the window is smaller than the beam 1 Hellma http www hellma worldwide com SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 55 eo gt 5 5 eo o D o P o s o 2 lt x co Part 8 Appendices Standard Semi micro and Microcuvettes Hellma omn 10 i gt Ir a T 10 00 mm Standard Semi micro Micro Hellma Cat No 100 104 105 004 104 002 108 002 105 Internal Dimensions 10 x 10 4x 10 4x 10 2x10 2x10 2x10 Fill Volume 4mL 1 4mL 600 uL 700 uL 500 uL 300 uL Ultra micro Cuvettes Hellma ie wast 30 00 mm Hellma Cat No 105 200 105 201 105 202 105 210 Fill Volume 180 uL 120 uL 70 uL 10 uL 56 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A
46. oduction 1 Description 1 1 Introduction SpectraMax and VersaMax microplate spectrophotometers provide rapid and sensitive measurements of a variety of analytes across a wide range of concentrations They measure the optical density OD of samples at selected wavelengths in a number of reading modes depending on the instrument model m i oO 7 oO EI Endpoint at a single point in time Kinetic over a specified period of time Spectral scan over a specified wavelength range The VersaMax and SpectraMax 190 read 96 well microplates The SpectraMax 340PC 84 and SpectraMax Plus 984 read both 96 and 384 well microplates The contents of the wells in a microplate can be mixed automatically by shaking before each read cycle which makes 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 be regulated if desired from 4 C above ambient to 45 C Detailed specifications of each model reader are reported in Section 7 Specifications 1 2 Components The main components of SpectraMax and VersaMax readers described in this manual are e Control panel e Microplate drawer 384 Cuvette chamber SpectraMax Plus only Back panel connections and power switch SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 1 Part 1 Descrip
47. oftMax Pro Express data as Absorbance or Transmittance Single wavelength reading of microplate Multiple wavelength up to six reading of microplate Kinetic and kinetic graphics of microplate Spectral scan 190 850 nm of microplate SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 45 zx 4 e Lr O 9 7 a S T o o o 2 7 Part 7 Specifications 46 MEASUREMENT TIME CALIBRATION OFF Read time endpoint standard read 96 wells in 9 seconds single wavelength 96 wells in 19 seconds dual wavelength 425 amp 650 nm Kinetic read intervals 96 wells 9 second minimum interval between readings single wavelength 1column 2 second minimum interval between readings single wavelength SCAN SPEED Microplate normal scan 33 K nm min 8 well strip Microplate speed scan 135 K nm min 8 well strip TEMPERATURE REGULATION Reading chamber Range Isothermal when temperature regulation is not enabled 4 C above ambient to 45 C when tempera ture regulation enabled Ambient tempera ture must be gt 20 C to achieve temperature regulation at 45 C Resolution 0 1 C 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
48. omix function permits automatic linear shaking along the long axis of the microplate at preset intervals thereby mixing of the contents within each well Auto mix must be selected before beginning a reading The actions associated with the Automix setting depend on the read mode chosen Endpoint mode Automix shakes the plate for a definable number of seconds and then reads at all selected wavelengths hn jas i ium N ae E amp A N Kinetic mode two types of Automix can be enabled Automix can shake the plate for a definable number of seconds before the initial reading and or for a definable number of seconds before each subsequent reading Use of Automix is strongly recommended for ELISAs and other solid phase enzyme mediated reactions to enhance accuracy 2 5 4 Computer Control SpectraMax and VersaMax readers are equipped with an 8 pin DIN RS 232 serial port through which the computer communicates with the instrument Different types of cables are available for connecting to different types of computers see Appendix 8 1 Cables SoftMax Pro software version 4 1 or greater is required to control SpectraMax and VersaMax instruments 16 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 3 1 Unpacking 3 Installation WARNING Always make sure the power switch on the instrument is in the OFF position and remove the power cord fro
49. p 77 K nm min 16 well strip TEMPERATURE REGULATION Reading chamber Isothermal when temperature regulation is not enabled Range 4 C above ambient to 45 C when tempera ture regulation enabled The ambient tem perature must be gt 20 C to achieve temperature regulation at 45 C Resolution 0 1 C 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 42 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 7 2 SpectraMax 340PC Specifications Temperature regulation 4 sensors Drift 0 2 C regulated Temperature regulation diagnostics Temperature regulation system is continu ously 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 between kinetic readings Plate mixing duration Selectable 1 to 999 seconds 3 second default COMPATIBILITY Microplates 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 plat
50. pre amp or exces sive stray light 313 reference gain check fail Power up check of the Reference ampli fier s gain out of tolerance 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 it faulty was commanded to in a 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 reasonable 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 can t find its photo interrupt 402 grating error Grating did not move to its photo inter rupt in a reasonable time or can t find its photo interrupt 403 stage error Stage did not move to its photo interrupt in a reasonable time or can t find its photo interrupt SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 33 o z an 7 EJ e Part 6 Troubleshooting 500 599 NVRAM ERRORS 500 NVRAM CRC corrupt The CRC for the NVRAM data is corrupt 501 NVRAM Grating cal data Grating calibration data is unreasonable bad 502 NVRAM Cuvette air cal Cuvette air calibration data is
51. r cord is securely plugged in to a functioning power outlet and to the receptacle at the rear of the SpectraMax or VersaMax If power failed while the instrument was already on Check that the power cord is not loose or disconnected and that power to the power outlet is functioning 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 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 25 m m gt o eo o rr D i Part 5 Maintenance U S and metric fuses are identical except for physical size They may be taped to the back of the instrument 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 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 To change fuses Switch power to the instrument off and then remove the power cord from the oudet and from the instrument power cord receptacle Remove the printer cable and computer cable if connected from the back of the instrument Turn the instrument around for easy access to the rear panel On the left hand side of the rear panel viewed from the back is the power switch fuse box and power cord receptacle
52. s the conventional basis for quantifying the unique absorbtivity properties of compounds in solution Quantitative analyses can SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 9 p mz 9 D 7 3 D to E Part 2 Principles of Operation be performed on the basis of extinction coefficients without standard curves e g NADH based enzyme assays When using a cuvette the pathlength is known and is independent of sample volume so absorbance is proportional to concentration In a microplate pathlength is dependent on the liquid volume so absorbance is pro portional to both the concentration and the pathlength of the sample Standard curves are often used to determine analyte concentrations in vertical beam photome try of unknowns yet errors can still arise from pipetting the samples and standards The PathCheck feature 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 correcting the microwell absorbance values is accurate to within 2 596 of the values obtained directly in a 1 cm cuvette i 5 i ium N i amp m N Horizontal Vertical light path light path Cuvette Microplate wells Figure 2 1 Cuvette and microwell light paths Reference measurements made by reading the cuvette Cuvette Reference Sp
53. so that the control panel now faces you Ensure no cables run beneath the instrument Leave at least three inches between the back of the instru ment and the nearest objects or surfaces to ensure proper ventilation and cooling 384 For the SpectraMax Plus f remove the tape from the cuvette door Turn on the power to the instrument wait for the microplate drawer to open and remove the tape and protective covering from the drawer subplate 18 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 4 1 Quick Overview 4 Operation This chapter contains operating information for SpectraMax and VersaMax Spectro photometers 4 1 Quick Overview If you are an experienced user of this instrument the following steps provide a quick reminder of the basic operating procedures required to perform an assay using a Spec traMax or VersaMax instrument 1 Turn on the power switch located on the back panel The microplate drawer opens automatically 2 Ifyou want to regulate the temperature inside the chamber touch the TEMP on off incubator key to turn the incubator on and bring the chamber to the default tem perature of 37 0 C The microplate drawer closes 3 Ifthe incubator is on the LCD shows the current temperature along with the tem perature set point To change the set point to any setting from ambient 44 to 45 C k 1 to z press the up or down arro
54. ssistance regarding warning messages contact your local Molecular Devices rep resentative 6 1 1 Error Code Classifications e mn 7 i EJ e Not all error messages are listed in this manual The errors are grouped in relationship to possible causes as follows SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 29 Part 6 Troubleshooting Table 6 1 SpectraMax and VersaMax error code ranges 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 assis tance of Technical Support 300 399 Instrument errors due to either a main board failure or other system failure Most of these errors require the assis tance of Technical Support 400 499 Errors caused by a motor motion failure Most of these errors require the assistance of Technical Support 500 599 Errors due to failure or improper initialization of the instru ments non volatile memory NVRAM All of these errors require the assistance of Technical Support Some errors shown in boldface in the following table are considered fatal in that if they are detected during power up the instrument aborts the power up sequence and displays FATAL ERROR on the LCD panel Check
55. the following list to see if there is something that you can do to change the condition of the instrument to prevent the fatal error e g closing the cuvette door during the power up sequence prevents errors 111 219 302 and 310 After correcting the problem leave the instrument on for about five minutes turn it off and then back on D i N 2 EI wo If you continue to get the fatal error message on power up record the error message number and contact Molecular Devices Technical Support or your local representa tive for assistance If the instrument is functioning normally when using SoftMax Pro no errors should be in the buffer except error number 100 30 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 RevA 6 1 Error Codes and Resolutions Table 6 2 Error codes error messages and notes about the errors ERROR MESSAGE NOTES ERROR CODE 100 199 UNRECOGNIZED COMMAND ERRORS SENT FROM THE COMPUTER 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 Instrument could not perform the give system busy command because it was busy doing another task 106 command invalid
56. ther microplate specifications apply to standard 96 well polystyrene flat bottom Technical specifications are subject to change without notice PHOTOMETRIC PERFORMANCE Wavelength range 190 850 nm Wavelength selection Monochromator tunable in 1 nm increments Wavelength bandwidth lt 2 0 nm full width half maximum Wavelength accuracy 1 0 nm across wavelength range Wavelength repeatability 0 2 nm across all optical channels Photometric range 0 to 4 000 OD 44 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 7 3 SpectraMax 190 Specifications Photometric resolution 0 001 OD Photometric accuracy linearity microplate 0 2 0 OD 190 850 nm 1 0 and 0 006 OD Photometric precision repeatability 0 2 0 OD 190 850 nm lt 1 0 and 0 003 OD Stray light lt 0 05 at 340 nm Photometric stabilization Instantaneous Photometric drift None continuous referencing of monochromatic input Calibration Automatic before first kinetic read and before every endpoint reading Optical alignment None required Light source Xenon flash lamp 5 Watts Average lamp lifetime 1 billion flashes Illumination Top down Photodetectors Silicon photodiode PHOTOMETRIC ANALYSIS MODES Standalone Single wavelength Absorbance read ing of the microplate Using S
57. tically reports absorbance values normalized to a 1 cm path length The table below shows results obtained with 75 pL to 300 pL yellow reagent Well Volume Pathlength Raw Absorbance HL cm Absorbance cm SB we 75 0 231 0 090 0 390 0 006 1 6 100 0 300 0 116 0 387 0 005 1 2 150 0 446 0 172 0 385 0 003 0 8 200 0 596 0 228 0 383 0 002 0 4 250 0 735 0 283 0 384 0 002 0 5 300 0 874 0 336 0 384 0 001 0 3 Absorbance in 1 cm cuvette 0 386 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 13 Part 2 Principles of Operation Optical pathlengths and raw absorbance values were directly proportional to well col umns After normalization to a 1 cm pathlength all absorbance values regardless of the volume in the wells were within 1 of the value obtained by measuring the same solution in a 1 cm cuvette 2 5 Functional Description The full power of a SpectraMax or VersaMax instrument can only be harnessed when the instrument is controlled using SoftMax Pro software running on a computer con nected to the instrument For a complete description of the modes of operation how to choose instrument settings etc refer to the SoftMax Pro User s Manual However some functionality is available directly on the instrument without having to use SoftMax Pro e Temperature control all models hon a i ium N ae E amp a
58. tion Cuvette Chamber Control Panel Back Panel 7 a 7 a Microplate Drawer Figure 1 1 SpectraMax Plus 9 components 1 2 1 The Control Panel The control panel consists of a 2 x 20 character LCD and eleven pressure sensitive membrane keys that can be used to control some functions of the instrument When you press a control panel key the instrument performs the associated action TEMP on off The TEMP on off key enables and disables the incubator e When the incubator is on the set temperature and actual temperature are shown on the front panel LCD display When the instrument is performing a kinetic or spectral scan the temperature keys on the front panel are disabled TEMP The TEMP keys allow you to enter a set point at which to regulate the microplate chamber temperature Pressing this key scrolls the temperature up or down starting at the previous temper ature setting or the default of 37 0 C if no setting had been made Pressing the up A or down V arrow once increments or decrements the dis played temperature by 0 1 C 2 SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 1 2 Components Pressing and holding either arrow increments or decrements the displayed temper ature by 1 C until it is released A not available on VersaMax Selects the wavelength to be used for reading the microplate manually m
59. to connect the instrument to the computer is a custom cable designed and built by Molecular Devices Please use the cable supplied by Molecular Devices or contact Molecular Devices for specific pin out requirements Male DB8 to Female DB9 custom cable made by Molecular Devices PN 9000 0149 USB Adapter iMac G4 and G5 Macintosh computers and many newer Windows computers do not have a serial port You can connect a serial cable between these computers and the instrument using a USB to serial adapter Molecular Devices has tested many third party serial to USB adapter cables and has found the Keyspan USA 19HS to be the most reliable It is the only one we recom mend and support Figure 8 1 Molecular Devices custom serial cable left and a serial to USB converter right eo gt 5 5 eo o D o SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 53 Part 8 Appendices 8 2 Accessories Description Part SpectraTest ABS1 Absorbance Validation Test Plate 0200 6117 Cuvette Absorbance Validation Kit 9000 0161 SpectraPlate Quartz UV transparent microplate R8024 Cable RS 232 8 pin DIN to 8 pin DIN instrument to pre G3 Macintosh 9000 0091 Cable RS 232 9 pin DIN to 8 pin DIN instrument to PC serial port 9000 0149 Power Cord US Canada Japan Mexico India 4400 0002 Power Cord EC1 Germany France Scandinavia Italy Korea 4400 00
60. ument If needed clean the microplate drawer using a cloth or sponge that has been damp ened with water Should fluids spill in the drawer area when the drawer is out they are directed to a tray at the bottom of the instrument from which they exit to the bench or counter beneath the instrument Wipe up any spills immediately Do not allow excess water or other fluids to drip inside the instrument 5 5 Cleaning the Fan Filter 5 6 The fan filter on the bottom of the instrument requires periodic cleaning The fre quency of cleaning depends on how dusty your particular lab is and could range from once a month to once every six months Turn power to the instrument OFF and then remove the power cord and cables from the back of the instrument Remove any plate or adapter from the instrument drawer Turn the instrument over so that it rests flat on the bench Pop the black fan cover off and remove the filter 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 Place the clean dry filter over the fan and replace the black cover Turn the instrument back over Reconnect the power cord and cables to the instrument 5 6 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 e Check to see whether the powe
61. w keys 4 Select the desired instrument settings read mode type of analysis template etc using SoftMax Pro software on the external computer 5 Ifyou are performing kinetic analysis add substrate at this time 6 Load the prepared microplate into the drawer being sure to match well A1 with the upper left hand corner of the drawer 7 Using SoftMax Pro start the reading 4 2 Preparing for a Reading 4 2 1 Turn the Instrument and Computer On The power switch is located on the back panel Press the rocker switch to the ON position The instrument automatically performs diagnostic checks to ensure that it is func tioning correctly Turn the computer on at this time also and start the SoftMax Pro software program SpectraMax and VersaMax Plate Readers Operator s Manual 0112 0126 Rev A 19 Part 4 Operation 4 2 2 Set the Temperature Optional To set the temperature within the microplate chamber you should turn on the incu bator first allowing enough time for the temperature to reach the set point before performing a reading When you first turn the instrument on up to 60 minutes may be required for the temperature within the chamber to reach the set point Turning on the incubator and choosing a temperature set point can be done using the software or the front panel of the instrument described here Temperature cannot be regulated at a set point that is lower than 4 C above the ambient temperature To ena

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