Synergy-HT Bio-Tek, Microplate Reader SOP

Primary Equipment:
Plate Reader
Created: February 23, 2010 by Tony Butterfield

Revised: February 23, 2010 by Tony Butterfield

University of Utah, Chemical Engineering

The Synergy-HT multi-mode microplate reader measures the absorbance, luminescence, and transmittance of many small volume samples, prepared in microplates. This SOP give a basic overview of its operation; a more detailed explanation of the plate reader�s features may be found in the manual.

  1. Turn on and login to the computer next to the plate reader (Figure 1). (username: biolab, password: biolab)
  2. Figure 1 width=
    Figure 1: Synergy-HT Bio-Tek Microplate Reader.
  3. Turn on the power to the plate reader. The switch is on the front of the device.
  4. The tray will open and present the microplate carrier.
  5. Start the program �KC4� from the computer�s desktop.
  6. Click the �Read� button.
  7. Select the detection method required for your assay (Absorbance at 660 nm for yeast cell concentration is common).
  8. Select Reading Type:
    1. End Point � Each well is given one value for each wavelength or filter at one point in time.
    2. Kinetic � The plate in this case is read multiple times at a particular wavelength at a given time interval (After picking this option, click the Kinetic button to set the interval).
    3. Spectrum � This mode only works with the Absorbance detection method. In this case a range of wavelengths are measured for each well.
  9. For Endpoint and Kinetic readings, select the wavelengths you wish to monitor (up to 6) from the dropdown list. For Spectrum reading, enter the wavelength start and stop points.
  10. Select the plate size (The standard size in the lab is the 8×12, 96 Well Plate).
  11. If you wish, you may also set the temperature within the chamber under the �temperature Control� heading.
  12. Click Ok once you have set all your settings.
  13. You may enter read descriptions in the Plate Reading dialogue box, or simply click �START READING�.
  14. The plate reader door will open, if it isn�t open already, and present the carrier. Insert your prepared sample plate and press �Ok�.
  15. The reading will then begin.
  16. After the reading is finished your data will appear in the Results window and the carrier will eject.
  17. To put the data into a format suitable for Excel, click File => Export. Select the data you wish to export (e.g. if one of your wavelengths was 570 nm, select �M#1 570� for export) and click �Add>>�. I you did a kinetic reading, you will need to select the read number min and max you wish to export under the �Kinetic readings� heading�, before you can add that data. Once you have selected all the desired data, click Ok and save it in a proper location. The resulting file is a comma-delimited text file.
  18. To take another reading, you must click the �New� button and begin the process again.
  19. When you are done, remove the plate, exit the software, close the tray , and turn the plate reader�s power off.