Estimation of the number of acceptor molecules surrounding a given donor in the (Forster resonance energy transfer) FRET system
An excitation-induced nonradiative energy transfer from one fluorophore (the donor) to another (the acceptor) occurs through long-range dipole-dipole coupling between the two fluorophores. The distance between the donor and acceptor is dependent on the FRET process. At sub-optical resolutions, fluorescence resonance energy transfer (FRET) is a very powerful method for detecting molecule interaction. You can figure out the FRET efficiency by taking the intensity of the donor in the presence of the acceptor and subtracting it from the intensity of the donor after photobleaching the acceptor. Then, you can compare this value to the intensity of the donor after bleaching. Acceptor photobleaching is one way of assessing FRET: the increase in donor fluorescence after full acceptor photobleaching is a measure of FRET efficiency.
Everything Saravanan said is correct of course. But I don't think it answers the question. I admit that I do not see a simple way to estimate the number of acceptors within FRET distance of a donor.
There is a user-friendly program called FRET-Calc, which offers a straightforward interface and is available for free. This program allows users to easily obtain a range of FRET parameters.
https://www.sciencedirect.com/science/article/abs/pii/S0010465523000607
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