Marshall Amplification Gemini EM User Manual
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Marshall Amplification Gemini EM User Manual

Marshall Amplification Gemini EM User Manual

Dual scanning microplate spectrofluorometer
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Gemini EM
Gemini XPS
Dual Scanning Microplate
Spectrofluorometer User Guide
Molecular Devices Corporation
1311 Orleans Drive Sunnyvale, California 94089
Part #0112-0128 Rev. A.
Gemini EM/XPS Dual-Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A

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  • Page 1 Gemini EM Gemini XPS Dual Scanning Microplate Spectrofluorometer User Guide Molecular Devices Corporation 1311 Orleans Drive Sunnyvale, California 94089 Part #0112-0128 Rev. A. Gemini EM/XPS Dual-Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 2 Molecular Devices Corporation, 1311 Orleans Drive, Sunnyvale, California, 94089, United States of America. Patents The Gemini EM, Gemini XPS, and methods have U.S. and International patents pending. 6,097,025, 6,232,608, 6,236,456, 6,313,471, 6,316,774, and 6,693,709. Gemini EM Patents 6,097,025, 6,232,608, 6,236,456, 6,313,471, and 6,316,774.
  • Page 3 Questions? Phone:1 (800) 635 5577 Fax:+1 (408) 747 3603 Web:www.moleculardevices.com Gemini EM/XPS Dual-Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 4 Gemini EM/XPS Dual-Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 5: Table Of Contents

    Changing the Fuses ..........29 Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 6 Gemini EM Specifications........

  • Page 7: Description

    1.1.4. WELL SCANNING Gemini EM and Gemini XPS can report a single point from the well center, or multiple data points from the bottom of large well tissue culture plates to provide high sensitivity for cell based assays.
  • Page 8 TOP AND BOTTOM READING OPTICS—GEMINI EM ONLY The top/bottom reading optical design of the Gemini EM allows for measurements for both solution and cell based assays. With the click of a button, the Gemini EM can be switched between top and bottom reading modes.

  • Page 9: Components

    When you press a control panel key, the Gemini performs the associated action. Figure 1.2: Control Panel. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 10 The adapter is required when using the SpectraTest FL validation plate to test the Gemini XPS and when testing the Gemini EM top read optics. To test the Gemini EM bottom reading performance, remove the purple adapter and then turn the validation plate upside down by rotating it from top to bottom so that column 1 remains on your left.
  • Page 11 UV range. If high sensitivity is required, it may be appropriate to use microplates that are designed to reduce background fluorescence. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 12 (PMT). This filter may be set automatically by the instrument or manually by the user. The PMT detects the emitted light and passes a quantitative signal to the instrument’s electronics that then send the data to the computer. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 13 The excitation beam is collimated by a mirror to a 1.0 mm diameter fiber before entering the sample in the microplate well. This focusing helps to prevent part of the beam from striking adjacent wells. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 14 Label: provides information about the Gemini, such as line voltage rating, cautionary information, serial number, etc. Record the serial number shown on this label for use when contacting Molecular Devices Technical Support. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 15: Principles Of Operation

    There is considerable overlap between the excitation and emission spectra (gray area) when a small Stokes shift is present. Excitation Emission maxim um maxim um Stokes shift Wavelength (nm) Figure 2.1: Excitation and emission spectra. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 16 The Gemini readers allow scanning of both excitation and emission wavelengths, using separate tunable monochromators. One benefit of being able to scan emission spectra is Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 17: Time Resolved Fluorescence

    The Gemini readers are microplate spectrofluorometers with photomultiplier tube detection. Some luminescence applications, such as gene reporter assays, may require a luminometer with photon counting detection for greater sensitivity. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 18: Functional Description

    For a complete description of the modes of operation, how to choose instrument settings, etc., refer to the SoftMax Pro User’s Manual. 2.4.1. READ MODES The Gemini EM and Gemini XPS can read in three modes: fluorescence, secondary luminescence, and secondary time resolved fluorescence. 2.4.2. READ TYPES Within each read mode, Gemini readers can perform four types of read: endpoint, kinetic, spectrum, and well scan.
  • Page 19 Temperature regulation and control of the microplate chamber is achieved through electric heaters, a fan, efficient insulation, and temperature sensors. The heaters are Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 20 The Gemini is 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 A, “Cables” and “Accessories”.) Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 21: Installation

    Insert the female end of the power cord into the power receptacle at the rear of the Gemini. Connect the male end to a grounded power outlet of the appropriate voltage. Gemini EM/XPS Dual Scanning Mocroplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 22: Installing The Drawer Adapter

    While pushing against the back edge of the adapter, lower the front of the adapter into the drawer. Figure 3.1: Adapter inserted in microplate drawer. Gemini EM/XPS Dual Scanning Mocroplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 23: Removing The Drawer Adapter

    3.4. REMOVING THE DRAWER ADAPTER If the adapter is in the drawer and you are either reading from the bottom(Gemini EM only) or using “high profile” (6 well, 12 well, 24 well, or 48 well) plates, you need to remove the adapter.
  • Page 24 3. Installation Gemini EM/XPS Dual Scanning Mocroplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 25: Operation

    The instrument automatically performs diagnostic checks to ensure that it is functioning correctly. Turn the computer on at this time also and start the SoftMax Pro software program. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 26: Read The Microplate

    If you return to the Gemini and find the drawer closed after a reading has finished, DRAWER press the key. When the drawer opens, you can remove the microplate. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 27: Optimizing Asays

    The excitation and emission wavelengths may be set in 1 nm increments within the range of the instrument. The Gemini EM reader allows excitation and emission wavelength scanning from 250 nm to 850 nm. The Gemini XPS reader allows excitation scanning from 250 nm to 850 nm and emission scanning from 360 nm to 850 nm.
  • Page 28 (Follow the plot to the left of the peak until the 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) Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 29 Select an emission cutoff filter that blocks as much of the residual excitation light as possible without unduly reducing the fluorescence signal. The cutoff wavelength choices are 325(Gemini EM only), 420, 435, 475, 495, 515, 530, 550, 570, 590, 610, 630, 665, or 695 nm. The cutoff value should be near the maximum emission wavelength (preferably between the excitation wavelength and the maximal emission wavelength) but at least 35 nm greater than the excitation wavelength.
  • Page 30 Figure 4.2 shows the effects of different cutoff filters on a scan of fluorescein where excitation was fixed at 485 nm and emission was scanned from 490 nm to 560 nm (buffer Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 31 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 For spectrum mode, the default is "manual" (no automatic cutoff ). Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 32 4. Operation Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 33: Maintenance

    If you have any problems using the Gemini EM or XPS Dual Scanning Microplate Spectrophotometer, in the U.S., contact our Technical Services group at 1 800 635 5577;...

  • Page 34: Cleaning

    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. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 35: Changing The Fuses

    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. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 36 Continue gently pulling the fuse box forward until it is free of the instrument. Figure 5.2: Removing the fuse box When removed, the fuse assembly will appear as shown in Figure 5.3. The holder inside contains two fuses. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 37 (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. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 38 5. Maintenance Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 39: Troubleshooting

    Maintenance procedures are described in the previous chapter. For problems with the Gemini EM or Gemini XPS 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.
  • Page 40 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 representative for assistance. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 41 200–299: FIRMWARE ERRORS 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. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 42 During a cuvette read, could be due to cuvette door being open. reference level saturation During a cuvette read, could be due to cuvette door being open. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 43 Grating did not move to its photo interrupt in a reasonable time, or can't find its photo interrupt. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 44 Stage offset data is unreasonable. 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. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 45: Specifications

    GENERAL PHOTOMETRIC PERFORMANCE Microplate formats 6, 12, 24, 48, 96, 384 Light source Xenon flash lamp (1 joule/flash) Average lamp lifetime 2 years normal operation (estimate) Detector Photomultiplier (R-3896) Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 46 13.5" (340 mm) × 16.5" (420 mm) × 16.5" (420 mm) Weight 35 lb (16 kg) Power consumption 500 VA maximum Line voltage and frequency 90–240 VAC, 50/60 Hz Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 47: Gemini Em Specifications

    GENERAL PHOTOMETRIC PERFORMANCE Microplate formats 6, 12, 24, 48, 96, 384 Light source Xenon flash lamp (1 joule/flash) Average lamp lifetime 2 years normal operation (estimate) Detector Photomultiplier (R-3896) Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 48 13.5" (340 mm) × 16.5" (420 mm) × 16.5" (420 mm) Weight 35 lb (16 kg) Power consumption 500 VA maximum Line voltage and frequency 90–240 VAC, 50/60 Hz Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 49: Appendix

    Keyspan USA 19HS (Molecular Devices, PN 9000 0938) to be the most reliable. It is the only one we recommend. Figure A.1: Molecular Devices' custom serial cable (left) and a serial-to-USB converter (right). Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 50: Accessories

    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 Adapter USB-Serial High-Speed (KeySpan adapter; instrument to USB-only 9000-0938 instrument) Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 51: Appendix

    COMMON WAVELENGTHS FOR FLUORESCENCE AND LUMINESCENCE Values in this table are based on the literature. You may want to scan your fluorochrome of interest in the Gemini EM or Gemini XPS to determine the optimal excitation and emission wavelengths for your application. B.1.1.

  • Page 52: Glossary

    Band pass filter that reduces the amount of extraneous lamp excitation light prior to the excitation monochromator. In endpoint reads and emission spectral scans, selection of excitation filter is automatic. In excitation spectral scans, the user has the choice of “no Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 53 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. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 54 The excitation lamp flashes and, after it is off, the delayed emission is collected for a set period of time before the lamp is flashed again. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

  • Page 55: System Diagrams And Dimensions

    B.3. System Diagrams and Dimensions B.3. SYSTEM DIAGRAMS AND DIMENSIONS Dimensions are shown in inches (millimeters). Figure B.1: Front view of Gemini EM/XPS. Figure B.2: Side view of Gemini EM/XPS. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...
  • Page 56 B. Appendix Figure B.3: Top view of Gemini EM/XPS. Gemini EM/XPS Dual Scanning Microplate Spectrofluorometer User Guide — 0112-0128 Rev. A...

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