Dear All,
I want to differentiate two fluorescence signals (SYBR green II and Fluorescein). Both of these have similar excitation, emission wavelength and color. So I am finding it difficult to differentiate between two. Looking for your suggestions and ideas to find the difference and differentiate.
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Dear Ismail,
there are at least two possibilities: measure the fluorescence decay function, and measure the polarization. Unfortunately to do this you need special equipment. And you cannot be sure to see a difference, even if it would be improbable that both dyes show the same values in all their properties.
Good success and best regards
Max
You need to differentiate first the conditions and processes you are using both the materials. Apparently, it seems you are using them in similar conditions though! It really need from you to spell out more on the problem!
Although the emission maxima of two dyes are similar, I think they will exhibit different spectral pattern in emission. You may calculate the FWHM of the emission spectra and try to match them with reported data. Otherwise you can do a lifetime measurement. Hope this helps.
One more advice: have a close look at your spectra, they are not identical: the maxima of the flu spectra differ by at least 10 nm, a little less in the excitation.
Regards
Max
Dear Max Lippitsch, Rajdeep and Riaz, Thanks for your valuable suggestions. I will try to find the difference in spectra to differentiate the fluorescence signals.
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