Molecular Devices SpectraMax M2 User Manual
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Molecular Devices SpectraMax M2 User Manual

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SpectraMax
M2/M2
user guide
®
A MULTI-DETECTION MICROPLATE READER WITH TWO-MODE CUVETTE PORT

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  • Page 1 SpectraMax M2/M2 user guide ® A MULTI-DETECTION MICROPLATE READER WITH TWO-MODE CUVETTE PORT...
  • Page 2 SpectraMax ® SpectraMax ® Multimode Plate Readers User Guide Molecular Devices Corporation 1311 Orleans Drive Sunnyvale, California 94089 Part #0112-0102 Rev. D. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 3 All other company and product names are 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 user guide is subject to change without notice.

  • Page 4: Table Of Contents

    Optimizing Fluorescence Assays ........28 SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 5 System Diagrams and Dimensions ........58 SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 6: Description

    200 nm and 1000 nm, any excitation wavelength between 250 nm and 850 nm, and any emission wavelength between 360 nm and 850 nm (SpectraMax M2) or 250 n and 850 nm (SpectraMax M2 ) for readings of both microplates and cuvettes.
  • Page 7 4°C above ambient to 45°C. 1.1.7. SUPPORTED PLATES Microplates having 6, 12, 24, 48, 96, and 384 wells can be used in the SpectraMax M2 and SpectraMax M2 . Top detection is available for fluorescence detection on the SpectraMax M2, while top and bottom reads are possible on the SpectraMax M2 .

  • Page 8: Components

    (enunciator blinks if not at set point). To see the microplate chamber temperature, you must use SoftMax Pro software. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...
  • Page 9 Pressing and holding either arrow increments or decrements the displayed temperature > by 1°C until it is released. You cannot set a temperature beyond the upper (45°C) or lower (15°C) instrument limits. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...
  • Page 10 (allowing for better robot integration—no springs or clips are used). The drawer remains in the reading chamber during read cycles. Figure 1.3: The microplate drawer. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...
  • Page 11 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. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...
  • Page 12 “reading” (clear) sides face left and right. Semi-micro and ultra-micro cuvettes can also be used with an adapter. Figure 1.5: Test tube cover. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...
  • Page 13 1. Description 1.2.5. THE BACK PANEL The following components are located on the back panel of the SpectraMax M2 and SpectraMax M2 Power switch: a rocker switch, labeled I/O (for on and off, respectively). > Power cord receptacle: plug the power cord in here.

  • Page 14: Principles Of Operation

    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- SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 15 Cuvette Reference and the Water Constant methods. The actual pathlength, d, of a solvent is found from the following equation: Sample OD – 1000 d cm -------------------------------------------------------------------- - SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 16 The cuvette values are stored temporarily and used in the PathCheck calculations for the microplate samples. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 17 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 solvent interference. However, you could correct for such interference by taking two cuvette measurements and SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 18: Fluorescence

    The figure below shows an example of excitation and emission spectra for a fluorophore. The excitation and emission bands are each fairly broad, with half- bandwidths of approximately 40 nm, and the wavelength difference between the SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 19 SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 20 SpectraMax M2 and SpectraMax M2 , refer to the section “Optimizing Fluorescence Assays” of the chapter “Operation” in this guide. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 21: Time-Resolved Fluorescence

    SoftMax Pro software running on a computer connected to the instrument. For a complete description of the modes of operation, how to choose instrument settings, etc. refer to the SoftMax Pro User Guide. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 22 1.0°C. The temperature uniformity within the microplate depends on its design and composition. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 23 Values reported are optical density, %Transmittance, relative fluorescence units (RFU), or relative luminescence units (RLU). SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 24 (Different types of cables are available for connecting to different types of computers (refer to the section “Cables” in the Appendix of this guide). SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 25 2. Principles of Operation SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 26: Installation

    Place the instrument on a level surface, away from direct sunlight, dust, drafts, vibra- tion, and moisture. Turn the instrument around so that the back of the instrument is facing you. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...

  • Page 27: Installing The Drawer Adapter

    While push- ing against the back edge of the adapter, lower the front of the adapter into the drawer. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...

  • Page 28: Removing The Drawer Adapter

    DRAWER in SoftMax Pro software. Remove the adapter plate. Figure 3.2: Microplate drawer without adapter. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...
  • Page 29 3. Installation SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers Operator’s Manual — 0112-0115 Rev. D...

  • Page 30: Operation

    If the incubator is on, the LCD shows the current temperature along with the temper- ature set point. To change the set point (to any setting from ambient +4° to 45°C), press the up or down arrow keys. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 31: Preparing For A Cuvette Or Microplate Reading

    Should you turn the incubator back on after a momentary shutdown, allow about ten minutes for the control algorithm to fully stabilize the chamber temperature. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 32: Read The Cuvette

    When the drawer opens, you can remove the microplate. DRAWER For more information about plate reading, please consult the SoftMax Pro User Guide. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 33: Optimizing Fluorescence Assays

    The temperature in the reading chamber may be adjusted from 4°C above ambient to 45°C. Note that assay optimization requires the use of a computer and SoftMax Pro software. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 34 RFU value falls to approximately 90% of the maximum, and then drop a line from the 90% point on the plot to the x-axis—see Figure 4.1.) SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 35 In SoftMax Pro, set up a third plate section for an emission scan as specified in step 3 above, except selecting Manual Cutoff Filter and setting the wavelength to that determined in step 4. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 36 695 nm cutoff or with no cutoff. Figure 4.2: Effects of cutoff filters on fluorescein. Emission was scanned from 490 to 560 nm; excitation was fixed at 485 nm. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 37 Endpoint and Kinetic Modes Wavelength (nm) Emission Wavelength (nm) None < 415 415–434 435–454 455–474 475–494 495–514 515–529 530–549 550–569 570–589 590–609 610–629 630–664 665–694 695–850 SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 38: Maintenance

    Damage caused by the use of incompatible or aggressive solvents is NOT covered by the instrument warranty. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 39: Moving A Spectramax M2 Or M2

    If required, clean the surfaces using a mild soap solution diluted with water or a glass > cleaner and then wipe with a damp cloth or sponge to remove any residue. Do not spray cleaner directly onto the instrument. > SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 40: Cleaning The Fan Filter

    Spare fuses (two U.S. and two metric) are shipped with the instrument. The U.S. and metric fuses are identical except for physical size. They may be taped to the back of the instrument. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 41 5. Maintenance 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).
  • Page 42 (see Figure 5.3). Figure 5.3: The fuse holder (with fuses), removed from the fuse box. 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.
  • Page 43 (toward the power receptacle). Press the fuse box into place, making sure the cover snaps closed. Reconnect the power cord to the instrument and to the wall outlet and reconnect other cables previously disconnected. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 44: Troubleshooting

    This action will not harm the instrument, but should only be taken if the first two options have failed to open the drawer. If you are still unable to open the drawer, contact your local Molecular Devices represen- tative.
  • Page 45 If you continue to get the fatal error message on power up, record the error message num- ber and contact Molecular Devices Technical Support or your local representative for assistance. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 46 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. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 47 During a cuvette read, could be due to cuvette door being open. plate air cal fail, low light Minimum signal/reference ratio not met during air calibration. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 48 Stage did not move to its photo interrupt in a reasonable time, or can't find its photo interrupt. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 49 NVRAM Battery Time to replace the NVRAM battery (U3). For all other error messages (codes not listed here), please contact your local Molecular Devices representative for assistance. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 50: Specifications

    Photometric accuracy/linearity, < ±1.0% and ±0.006 OD 0–2.0 OD Photometric precision (repeatability), < ±1.0% and ±0.003 OD 0–2.0 OD ≤ 0.05% at 230 nm Stray light SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 51 96-well black plates: auto gain circuitry System validation Self-calibrating with built-in fluorescence cali- brators Light source Xenon flash lamp (1 joule/flash) Average lamp lifetime 2 years normal operation Detector Photomultiplier (R3896) SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 52 Microplate read time (endpoint), • 96 wells in 2:07 minutes (single Standard read with PathCheck wavelength, absorbance) • 384 wells in 7:19 minutes (single wavelength, absorbance) SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 53 Flat-bottom microplates with isolated wells and lid Control Front panel reports cuvette chamber temper- ature only (temperature for microplate cham- ber reported in SoftMax Pro) AUTOMIX WITH SOFTMAX PRO SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 54 8-pin DIN RS-232 serial (double shielding required) Printer interface Parallel 25-pin to Centronics (double shielding required) Microplates supported All 6- to 384-well and strip-well microplates, including lids SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 55 8.6" (220 mm) x 22.8" (580 mm) x 15" (380 mm) Weight 35 lbs. (15.75 kg) Power consumption < 250 W Line voltage and frequency 90–250 V autoranging, 50/60 Hz SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 56: Appendix

    Keyspan USA-19HS (Molecular Devices, PN 9000-0938) to be the most reli- able. It is the only one we recommend. Figure A.1: Molecular Devices' custom serial cable (left) and a serial-to-USB converter (right). SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 57: Accessories

    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. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 58 (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 M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 59: Common Wavelengths For Fluorescence And Luminescence

    Values in this table are based on the literature. You must scan your fluorochrome of inter- est in the SpectraMax M2 or SpectraMax M2 to determine the optimal excitation and emission wavelengths for your application. Excitation and emission wavelengths listed by SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 60: Fluorescence

    Attophos /Attofluor FITC Ethidium Homodmer (DNA) TRITC, Ethidium Bromide Texas Red TAMRA Tryptophan La Jolla Blue A.6. TIME-RESOLVED FLUORESCENCE Fluorophore Excitation Wavelength (nm) Emission Wavelength (nm) Eu-Chelate SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 61: Luminescence

    (or no excitation in the case of luminescence). The default value reported for each well is the wavelength of maximum fluorescence or luminescence. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 62 The emission of light by processes that derive energy from essentially non-thermal changes, the motion of subatomic particles, or the excitation of an atomic system by radi- ation. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...

  • Page 63: System Diagrams And Dimensions

    A.9. SYSTEM DIAGRAMS AND DIMENSIONS Dimensions are shown in inches (millimeters). Figure A.2: Front view of SpectraMax M2. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 64 A.9. System Diagrams and Dimensions Figure A.3: Side view of SpectraMax M2. Figure A.4: Top view of SpectraMax M2. SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 65 A. Appendix SpectraMax M2 & SpectraMax M2e Multi-mode Plate Readers User Guide — 0112-0102 Rev. D...
  • Page 66 Fax +44-118-944-8001 Germany www.moleculardevices.com Molecular Devices GmbH Tel. +49-89-9605-880 ©2006 Molecular Devices Corporation. Printed in U.S.A. Fax +49-89-9620-2345 SpectraMax is a registered trademark of Molecular Devices Corporation . All other trademarks are the property of their respective owners. Specifications subject to change without notice.

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