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- W3122557777 abstract "A new method of flow cytometric dual-laser FRET calibration is presented based on an information theoretical description of the FRET signals. This method belongs to the so called two-channel or ratiometric FRET methods, whose crucial and common element is the determination of a calibration factor called α (alpha- or G-factor), to place the detected FRET signal on the absolute scale of efficiency. Generally α is determined by extra cell samples labeled with only donor and only acceptor, where the donor-acceptor ratio is known. Alternatively, when the donor-acceptor ratio is not known, standard donor-acceptor FRET pairs of known FRET efficiency can also be used for determining α. In our approach, there is no need for extra information presented by the known acceptor-to-donor ratio or the standard FRET efficiency. Our approach proceeds solely on the signals of the interested FRET-pair applying only a single FRET sample, not necessitating extra ones. The missing information of extra cell samples is replaced by an information theoretic balance equation written for the actual FRET sample. Our method has been proven to be robust, inasmuch as both donor and acceptor appear in adequate levels in the FRET sample used for finding α." @default.
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- W3122557777 date "2021-03-01" @default.
- W3122557777 modified "2023-10-16" @default.
- W3122557777 title "Information theoretic FRET calibration on the cell surface" @default.
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- W3122557777 doi "https://doi.org/10.1016/j.jphotochem.2021.113144" @default.
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