Hello everyone,
In my lab we have an epifluorescence microscope. Since we purchased the filter cubes from Chroma, I often go to the Chroma Spectra Viewer (https://www.chroma.com/spectra-viewer) to help me visually line up my available filter sets with my fluorophores of interest. They also offer data on OD and Transmission % for the fluorophores and filter sets and I often wondered if there was more I could do with that raw data. What additional mathematical analyses could I perform using the filter set and fluorophore spectra data that might help in the choosing the best fluorophore to match a filter set (quantitatively). For the same lamp intensity and exposure time, is there a way to predict which filter set/fluorophore combination will theoretically give me the strongest signal ? I want to say that the integral of the spectra could be used for something, but I'm not sure what.
Your spectra have asymmetric shape = a sum of components. Each component has specific parameters, e.g., max wavelength. Simple deconvolution of spectra (e.g., the second derivative) may help you to establish relationships between chemical structure of fluophores and internal structure of spectra. There are many materials in fluorescent spectroscopy which concerned your problem.
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