Since my samples are only 200 ul they evaporate quickly if left at 37C in the plate reader. I tried PerkinElmer TopSeal-A Plus membranes to seal the wells and hold in the moisture. They say they are heat resistant and are clear and should not effect fluorescence, but my controls like pure water and PBS are very noisy with the membrane.
I want to know if I could use the TopSeal-A Black membranes and use bottom reads to minimize the scattering from the condensation on the membrane. But does bottom reading on Spectra Max M5 get excitation pulsed and emission reading from the bottom?
I don't know the answer but from a purely physical perspective, condensation on the membrane, to the extent that it occurs, will deplete the residual solution volume in the well. Could you prevent the condensation by ensuring a small, positive temperature differential at the top? The if cooling is needed before measurement could it be from the bottom?
The TopSeal Black membranes have worked very well for me. Using black clear bottom 96 well plates with the black TopSeal membrane keeps my samples and has minor background.
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