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Information theoretic FRET calibration on the cell surface
Journal of Photochemistry and Photobiology A: Chemistry ( IF 4.3 ) Pub Date : 2021-01-15 , DOI: 10.1016/j.jphotochem.2021.113144
László Bene , Miklós Bagdány , Tamás Ungvári , László Damjanovich

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.



中文翻译:

细胞表面信息理论FRET校准

基于FRET信号的信息理论描述,提出了一种新的流式细胞术双激光FRET标定方法。此方法属于所谓的两通道或比率FRET方法,其关键和共同的要素是确定称为α(α或G因子)的校准因子,以将检测到的FRET信号置于绝对效率范围内。 。通常,α是由额外的仅标有供体和受体的细胞样品确定的,其中供体与受体的比例是已知的。可替代地,当未知供体-受体比时,已知FRET效率的标准供体-受体FRET对也可以用于确定α。在我们的方法中,不需要通过已知的受主与受主比率或标准FRET效率提供额外的信息。我们的方法仅对感兴趣的FRET对的信号进行处理,而仅应用单个FRET样本,而不需要额外的样本。多余的细胞样本缺少的信息将替换为针对实际FRET样本编写的信息理论平衡方程。我们的方法已被证明是可靠的,因为供体和受体在用于发现α的FRET样品中均以足够的水平出现。

更新日期:2021-01-22
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