I'm checking fluorescence of umbliferon and has set water as blank in two wells. The two wells are showing same reading but one is +ve and another is -ve.
Also, the fluorescence can't be negative. So, please tell me any explanation or the mistake I'm doing.
Hi Deepchandra Joshi
I’m assumibg you are using a plate reader for your readings? With the given information, this could be due to many variables. However, did you try to repeat the reading using a different 96 well plate first? Do you take top reading or bottom reading for the emission? How different your values for two controls?
Chathura S. Abeywickrama I didn't try it with any other 96 well plate reader. I got exactly the same values, only one is +ve and another is -ve. I am using the bottom reading for the emission.
If I neglect the blank reading I got the correct readings for other samples.
It would be preferable to perform top reading in a solid black plate to eliminate well-to-well crosstalk and fluorescence from the plastic.
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