I am trying to do fluorescnce polarisation anisotropy with Bocilin and proteins. How will I fix the aoptimum gain and z position value for the plate reader. I am using the Tecan plate reader and using 384 well plates
Hi Debasmita,
Are you working with a Tecan Spark plate reader?
If yes please follow these steps:
1) Select the correct plate and the wells that you want to measure
2) Add in your protocol in Method Editor the Fluorescence Polarization strip
3) At the bottom of the strip, click on "Show advanced settings"
4) Check that Gain is set to "Optimal"
5) Select for Z-position the setting "Calculated from well" and choose a well with a strong fluorescent signal (important to avoid focusing on noise)
6) Run your protocol
At the end of the measurement, an excel file will open where you will find the measurement results. In this same sheet are shown the different measurement parameters, included the optimized ones. For example, the gain will be optimized at a certain value (it should be lower than 150), and this will be written in the excel report. Same for the Z-position.
If you want to repeat this assays in the future and compare it with this run, go back to the Method Editor and set Gain and Z-position to "Manual". Now insert the respective optimized parameters that you can find in the Excel result sheet, and save the protocol.
I hope this answered your question!
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