What is the main difference between dark or clear bottom plates to measure fluorescence?
There are different types of plates available and you should choose to match the device configuration. If you are using a system which allows fluorescence excitation and detection from the top then a dark bottom plate would be great. If thats not the case and the sample is excited from top while detection is from the bottom you would need a dark plate that has a transparent bottom. Usually dark plates are prefered for fluorescence as there is less scattering from the well walls and allows better collection of emitted light .If you have lot of samples and dont leave space between wells in a clear plate, it prevents crosstalk. If your samples are intense and you can load them a few wells apart you could still get a decent result from clear plates after running blank to evaluate background fluorescence in an empty well or with diluent but no fluorophore
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There are different types of plates available and you should choose to match the device configuration. If you are using a system which allows fluorescence excitation and detection from the top then a dark bottom plate would be great. If thats not the case and the sample is excited from top while detection is from the bottom you would need a dark plate that has a transparent bottom. Usually dark plates are prefered for fluorescence as there is less scattering from the well walls and allows better collection of emitted light .If you have lot of samples and dont leave space between wells in a clear plate, it prevents crosstalk. If your samples are intense and you can load them a few wells apart you could still get a decent result from clear plates after running blank to evaluate background fluorescence in an empty well or with diluent but no fluorophore
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Well said by Nagarajan. Otherwise it doesn't matter what colour the plate is. Just ensure there is no light scattering if you are using top excitation and detection flourimeter. Although for my caspase 3 activity, manufacturer instruction indicated black plate, but our Central lab had a flourimeter with white plate plus my black plate didn't fit in the plate drive. Your results won't be affected, I hope.
Thank you so much both of you, Sounderya Nagarajan and Ajayi. Very helpful
I think the black flat bottom plate should be used. However, it might depend on the machine config. Hope that helps.
Machine configuration is the main thing, whether it measures from upward or downward direction.
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