Computer program for analyzing donor photobleaching FRET image series

The photobleaching fluorescence resonance energy transfer (pbFRET) technique is a spectroscopic method to measure proximity relations between fluorescently labeled macromolecules using digital imaging microscopy. To calculate the energy transfer values one has to determine the bleaching time constants in pixel‐by‐pixel fashion from the image series recorded on the donor‐only and donor and acceptor double‐labeled samples. Because of the large number of pixels and the time‐consuming calculations, this procedure should be assisted by powerful image data processing software. There is no commercially available software that is able to fulfill these requirements.

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