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Summary of Contents for Molecular Devices SpectraMax M2
- Page 1 SpectraMax® M2 SpectraMax® M2e Multi-Mode Microplate Readers User Guide 0112-0102 E October 2018...
- Page 2 For research use only. Not for use in diagnostic procedures. The trademarks mentioned herein are the property of Molecular Devices, LLC or their respective owners. These trademarks may not be used in any type of promotion or advertising without the prior written permission of Molecular Devices, LLC.
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Page 3: Table Of Contents
Contents Safety Information Warnings, Cautions, Notes, and Tips Chapter 1: Introduction Computer Integration Supported Plates and Cuvettes Shake Temperature Regulation Chapter 2: Setting Up the Instrument Connecting Instrument Cables Chapter 3: Getting Started Control Panel Cuvette Chamber Plate Drawer Plate Adapter Chapter 4: Operating the Instrument Temperature Settings Wavelength Settings... - Page 4 SpectraMax M2 and M2e Microplate Reader User Guide Chapter 7: Troubleshooting Opening the Drawer Manually Obtaining Support Appendix A: Specifications System Diagrams and Dimensions Appendix B: Accessories Appendix C: Electromagnetic Compatibility 0112-0102 E...
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Page 5: Safety Information
Follow the related safety information. The following user attention statements might be displayed in the text of Molecular Devices user documentation. Each statement implies the amount of observation or recommended procedure. - Page 6 For products under the requirement of the WEEE directive, contact your dealer or local Molecular Devices office for the procedures to facilitate the proper collection, treatment, recovery, recycling, and safe disposal of the device. - - California proposition 65 requires businesses to provide warnings to Californians about significant exposures to chemicals that cause cancer, birth defects, or other reproductive harm.
- Page 7 Safety Information To protect against fire hazard, replace the fuses only with the same type and rating as the original factory installed fuses. To ensure sufficient ventilation and provide access to disconnect power from the instrument, maintain a 20 cm to 30 cm (7.9 in. to 11.8 in.) gap between the rear of the instrument and the wall.
- Page 8 SpectraMax M2 and M2e Microplate Reader User Guide Moving Parts Safety The instrument contains moving parts that can cause injury. Under normal conditions, the instrument is designed to protect you from these moving parts. WARNING! If the instrument is used in a manner not specified by Molecular Devices, the protection provided by the equipment might be impaired.
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Page 9: Chapter 1: Introduction
Dual monochromators enable you to select absorbance wavelengths between 200 nm and 1000 nm, excitation wavelengths between 250 nm and 850 nm, and emission wavelength between: 360 nm and 850 nm (SpectraMax M2). 250 nm and 850 nm (SpectraMax M2e). 0112-0102 E... -
Page 10: Computer Integration
Endpoint, Kinetic, Spectrum, and Well Scan read types are available for the Absorbance and Fluorescence read modes with any plate model. Top reads are available for the Fluorescence read mode for the SpectraMax M2. Top and bottom reads are available for the Fluorescence read mode for the SpectraMax M2e. -
Page 11: Shake
Endpoint, Kinetic, and Spectrum read types are available for the Absorbance and Fluorescence read modes with cuvettes. Note: You can use a 12 x 75 mm test tube with a test tube cover. You can order the test tube cover from Molecular Devices. See Accessories on page Shake The software enables you to define settings to mix of the contents of the wells by shaking the plate linearly along the long axis at preset intervals. -
Page 12: Chapter 2: Setting Up The Instrument
Chapter 2: Setting Up the Instrument Before you unpack and set up the instrument, prepare a dry, flat work area that is away from direct sunlight, dust, drafts, vibration, and moisture. The work area must have sufficient space for the instrument, host computer, and required cables. To provide access for disconnecting power from the instrument, maintain a 20 cm to 30 cm (7.9 in. -
Page 13: Connecting Instrument Cables
SpectraMax M2 and M2e Microplate Reader User Guide Connecting Instrument Cables The power cord and USB cable connect to the ports on the rear of the instrument. Part Number Description 1-2100-0548 USB computer connection cable, 3 meter (9.8 foot) 4400-0002 Power cord, 1 meter (3.3 foot) See... -
Page 14: Chapter 3: Getting Started
Chapter 3: Getting Started Now that you have installed the SoftMax Pro Software software on the computer, removed the tape from the cuvette port, and connected the cables, it is time to get started. Turn on the instrument power switch located on the rear of the instrument. Wait for the instrument to complete its diagnostic check and the plate drawer opens. -
Page 15: Control Panel
SpectraMax M2 and M2e Microplate Reader User Guide Control Panel The instrument control panel works in conjunction with the SoftMax Pro Software to open the plate drawer, adjust the temperature, and to set the wavelengths for cuvette reads. See the SoftMax Pro Data Acquisition and Analysis Software User Guide. - Page 16 Chapter 3: Getting Started Temp The Temp p and q buttons enable you to enter the set point at which to regulate the temperature inside the cuvette chamber and the plate chamber. The temperature for the instrument chambers is isothermal at ambient temperature as well as at elevated temperatures.
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Page 17: Cuvette Chamber
SpectraMax M2 and M2e Microplate Reader User Guide Cuvette Chamber The cuvette chamber is located to the right of the control panel. Lift up the lid over the chamber to insert or remove a cuvette. The chamber contains springs that position the cuvette in the proper alignment for a read. -
Page 18: Plate Drawer
Chapter 3: Getting Started Plate Drawer The plate drawer is located on the right side of the instrument and slides in and out of the plate chamber. An internal latch positions the plate on the drawer as it closes. You can use 6, 12, 24, 48, 96, and 384-well plates that conform to ANSI/SBS standard microplate footprint and dimensions in the instrument. -
Page 19: Plate Adapter
SpectraMax M2 and M2e Microplate Reader User Guide Plate Adapter Before you read standard 96-well or 384-well plates from the top, you must insert the plate adapter in the plate drawer. Before you do a bottom read, or use high profile (6-well, 12-well, 24-well, or 48-well) plates, you must remove the plate adapter from the plate drawer. -
Page 20: Chapter 4: Operating The Instrument
Chapter 4: Operating the Instrument You do the same basic steps to operate the instrument when you read a plate or you read a cuvette. Note: For detailed software instructions, see the SoftMax Pro Data Acquisition and Analysis Software User Guide or the application help. Temperature Settings When you use the incubator to set the temperature in the cuvette chamber and the plate chamber, you should wait at least 60 minutes for the chamber temperature to reach the set... -
Page 21: Wavelength Settings
Common Wavelengths The SpectraMax M2 and SpectraMax M2e enable you to scan the fluorophore of interest to determine the optimal excitation and emission wavelengths. Excitation and emission wavelengths listed by fluorophore manufacturers are determined in methanol and do not reflect actual values because of changes in pH, salt content, and so on. -
Page 22: Read A Plate
Chapter 4: Operating the Instrument Fluorescence Fluorophores Wavelengths Fluorophore Excitation Wavelength (nm) Emission Wavelength (nm) HPPA 4-MeU, NADH, NADPH Biotinidinase Green Fluorescent Protein Attophos /Attofluor FITC Ethidium Homodimer (DNA) TRITC, Ethidium Bromide Texas Red TAMRA Tryptophan Read a Plate CAUTION! Before you read standard 96-well or 384-well plates from the top, you must insert the plate adapter into the plate drawer. -
Page 23: Read A Cuvette
SpectraMax M2 and M2e Microplate Reader User Guide 5. Place the plate onto the plate drawer, matching well A1 with position A1 in the drawer. Note the following: If you read 6, 12, 24, or 48-well plates, make sure that the plate is flat against the drawer bottom. -
Page 24: Chapter 5: Read Modes And Read Types
Chapter 5: Read Modes and Read Types Absorbance Read Mode The instrument uses the Absorbance (ABS) read mode to measure the Optical Density (OD) of the sample solutions. Absorbance is the quantity of light absorbed by a solution. To measure absorbance accurately, it is necessary to eliminate light scatter. - Page 25 SpectraMax M2 and M2e Microplate Reader User Guide The factor of two comes from the fact that %T is expressed as a percent of the transmitted light and log (100) = 2. When in %Transmittance analysis mode, the instrument converts the raw OD values reported by the instrument to %Transmittance using the above formula.
- Page 26 Chapter 5: Read Modes and Read Types In a plate, pathlength is dependent on the liquid volume, so absorbance is proportional to both the concentration and the pathlength of the sample. Standard curves are often used to determine analyte concentrations in vertical-beam photometry of unknowns, yet errors can still occur from pipetting the samples and standards.
- Page 27 SpectraMax M2 and M2e Microplate Reader User Guide Place a standard 1 cm cuvette that contains the aqueous/solvent mixture you use for the plate samples into the cuvette port. The cuvette must be in place when you read the plate.
- Page 28 Chapter 5: Read Modes and Read Types 1. Fill a clean plate with water. 2. Read at the wavelengths you will use for the samples. The average OD value is the Plate Background OD. If you intend to read your samples at more than one wavelength, there should be a corresponding number of Plate Background OD values for each wavelength.
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Page 29: Fluorescence Intensity Read Mode
SpectraMax M2 and M2e Microplate Reader User Guide Fluorescence Intensity Read Mode Fluorescence occurs when absorbed light is re-radiated at a longer wavelength. In the Fluorescence Intensity (FL) read mode, the instrument measures the intensity of the re- radiated light and expresses the result in Relative Fluorescence Units (RFU). - Page 30 Chapter 5: Read Modes and Read Types Tip: If the Stokes shift is small, you should choose an excitation wavelength that is as far away from the emission maximum as possible while still able to stimulate the fluorophore so that less of the excited light overlaps the emission spectrum, which permits better selection and quantitation of the emitted light.
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Page 31: Luminescence
Optimizing Luminescence Read Mode You can do top Luminescence reads with the SpectraMax M2. You can do top or bottom Luminescence reads with the SpectraMax M2e. You should use solid white plates or white plates with clear bottoms for Luminescence assays. -
Page 32: Time-Resolved Fluorescence Read Mode
Chapter 5: Read Modes and Read Types You do not designate luminescence read times based on multiple reads per well, but rather by choosing the total integration time that is between 1 ms and 1,500 ms. Typical luminescence assays require between 500 ms and 1,000 ms for integration time. If wells incubate for a long period of time, then you should use the Automix function to mix the wells before the read. -
Page 33: Read Types
Optimizing Time-Resolved Fluorescence Read Mode The SpectraMax M2 enables you to read Time-Resolved Fluorescence assays from the top of a plate. The SpectraMax M2e enables you to read Time-Resolved Fluorescence assays from either the top or the bottom of a plate. You should use solid white plates for top Time- Resolved Fluorescence reads, and white plates with clear bottoms for bottom reads. - Page 34 Chapter 5: Read Modes and Read Types The kinetic read type can collect data points in time intervals of seconds, minutes, or hours (up to 99 hours). Kinetic analysis has many advantages to determine the relative activity of an enzyme in different types of plate assays, including ELISAs and the purification and characterization of enzymes and enzyme conjugates.
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Page 35: Chapter 6: Maintenance
Chapter 6: Maintenance Perform only the maintenance tasks described in this guide. Contact a Molecular Devices service engineer to inspect and perform a preventive maintenance service on the instrument each year. See Obtaining Support on page Before you operate the instrument or perform maintenance operations, make sure you are familiar with the safety information in this guide. -
Page 36: Replacing Fuses
SpectraMax M2 and M2e Microplate Reader User Guide Always turn the power switch off and disconnect the power cord from the main power source before using liquids to clean the instrument. Periodically clean the outside surfaces of the instrument using a cloth or sponge that has... - Page 37 Chapter 6: Maintenance If these checks fail to remedy the loss of power, replace the fuses. You can obtain replacement fuses from Molecular Devices. CAUTION! Do not touch or loosen screws or parts other than those specifically designated in the instructions. Doing so could cause misalignment and possibly void the warranty.
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Page 38: Before You Move The Instrument
SpectraMax M2 and M2e Microplate Reader User Guide Before You Move the Instrument Before you move the instrument, make sure that the new location is a dry, flat work area that has sufficient space for the instrument, host computer, and required cables. -
Page 39: Chapter 7: Troubleshooting
#100, no errors should be in the buffer. This guide does not list all possible error codes. For assistance with all other error messages (codes not listed here), please contact Molecular Devices Technical Support or your local representative. - Page 40 SpectraMax M2 and M2e Microplate Reader User Guide Note: If an error occurs while the plate drawer is closed, and you need to remove a plate, see Opening the Drawer Manually on page Table 7-2: Error Codes 100-199: Unrecognized Command Errors Sent From the Computer...
- Page 41 Chapter 7: Troubleshooting Table 7-3: Error Codes 200-299: Firmware Errors Error Error Message Notes Code assert failed Firmware error. bad error number Firmware error. receive queue overflow Caused by external device sending too much data over serial port and ignoring flow control. serial port parity error Parity bit error detected with incoming serial data.
- Page 42 SpectraMax M2 and M2e Microplate Reader User Guide Table 7-4: Error Codes 300-399: Hardware Errors Error Error Message Notes Code thermistor Unable to read a reasonable thermistor value. Thermistor faulty or faulty disconnected, Main board problem, or ambient temperature out of range.
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Page 43: Opening The Drawer Manually
(do not force the drawer) and remove the plate. This action will not harm the instrument, but should only be taken if the first two options fail to open the drawer. If you are still unable to open the drawer, contact your local Molecular Devices representative. -
Page 44: Obtaining Support
SpectraMax M2 and M2e Microplate Reader User Guide Obtaining Support Molecular Devices is a leading worldwide manufacturer and distributor of analytical instrumentation, software, and reagents. We are committed to the quality of our products and to fully supporting our customers with the highest level of technical service. -
Page 45: Appendix A: Specifications
100 μL and 300 μL. Note: Technical specifications are subject to change without notice. Molecular Devices provides validation documentation for software and hardware, as well as absorbance, fluorescence, and luminescence detection test tools with its SpectraTest®... - Page 46 SpectraMax M2 and M2e Microplate Reader User Guide Table A-1: Absorbance Photometric Performance Item Description Wavelength range 200-1000 nm Wavelength selection Monochromator, tunable in 1 nm increments Wavelength bandwidth ≤ 4.0 nm full width half maximum Wavelength accuracy ±2.0 nm across wavelength range Wavelength repeatability ±0.2 nm...
- Page 47 Appendix A: Specifications Table A-2: Fluorescence Intensity Performance Item Description Sensitivity Top Read (M2 and M2e) 3.0 fmol/well in 200 µL FITC (96 well microplate and 15 pM) 3.0 fmol/well in 75 µL FITC (384 well microplate and 40 pM) Bottom Read (M2e only) 5.0 fmol/well in 200 µL FITC (96 well microplate and 25 pM) 5.0 fmol/well in 75 µL FITC (384 well microplate and 67 pM)
- Page 48 SpectraMax M2 and M2e Microplate Reader User Guide Table A-3: Time-Resolved Fluorescence Performance Item Description Sensitivity 0.5 fmol/well Eu-Ch Wavelength range 250-850 nm Wavelength selection Monochromators, tunable in 1 nm increments Bandwidth (excitation, emission) 9 nm, 9 nm Precision data collection 1–100 flashes;...
- Page 49 Appendix A: Specifications Table A-6: Measurement Time (Calibration Off) Item Description Microplate read time (endpoint), 96 wells in 24 seconds (single wavelength, Standard read absorbance) 96 wells in 15 seconds (single wavelength, fluorescence intensity) 384 wells in 1:57 minutes (single wavelength, absorbance) 384 wells in 45 seconds (single wavelength, fluorescence intensity)
- Page 50 SpectraMax M2 and M2e Microplate Reader User Guide Table A-8: Temperature Regulation Item Description Reading chamber Isothermal when temperature regulation is not enabled Range 4°C above ambient to 45°C when temperature regulation enabled. Note: To achieve temperature regulation at 45°C, the ambient temperature must be >...
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Page 51: System Diagrams And Dimensions
Appendix A: Specifications System Diagrams and Dimensions In the following drawings, the dimensions are show in centimeters and inches. Front View with Dimensions Side View with Dimensions 0112-0102 E... - Page 52 SpectraMax M2 and M2e Microplate Reader User Guide Top View with Dimensions 0112-0102 E...
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Page 53: Appendix B: Accessories
Appendix B: Accessories This appendix describes supported accessories for the instrument. Supported Plates and Cuvettes The software supports the following plates: 96-well Standard, 96 Costar, 96 Greiner Black, 96 Bottom Offset, 96 Falcon, 96 BD Optilux/Biocoat, 96 BD Fluoroblok MW Insert, 96 Corning Half Area, 96 MD HE PS 384-well Standard, 384 Costar, 384 Greiner, 384 Falcon, 384 Corning, 384 MD HE PS 48 Costar 24 Costar... - Page 54 SpectraTest Microplate Reader Validation Packages Technical specifications are subject to change without notice. Molecular Devices provides validation documentation for software and hardware, as well as absorbance, fluorescence, and luminescence detection test tools with its SpectraTest® solutions. The SpectraTest line of microplate reader validation packages provide automated and comprehensive validation of a microplate reader's optical performance.
- Page 55 Appendix B: Accessories Additional Accessories The following accessories and international power cords are compatible with the instrument. Accessories Item Name Part Number SpectraPlate—Quartz UV-transparent R8024 plate Fuse, 4-amp Time Delay 4601-0013 Fuse, 4-amp (5 x 20 mm) Time Delay 4601-0014 Test Tube Cover 2300-0277 Power cord, USA/Canada...
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Page 56: Appendix C: Electromagnetic Compatibility
Appendix C: Electromagnetic Compatibility Regulatory for Canada (ICES/NMB-001:2006) This ISM device complies with Canadian ICES-001. Cet appareil ISM est confomre à la norme NMB-001 du Canada. ISM Equipment Classification (Group 1, Class A) This equipment is designated as scientific equipment for laboratory use that intentionally generate and/or use conductively coupled radio-frequency energy for internal functioning, and are suitable for use in all establishments, other than domestic and those directly connected to a low voltage power supply network which supply buildings used for domestic... - Page 57 The trademarks used herein are the property of Molecular Devices, LLC or their respective owners. ©2018 Molecular Devices, LLC. Specifications subject to change without notice. Patents: www.moleculardevices.com/productpatents All rights reserved. FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES...
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