Improving FRET efficiency measurement in confocal microscopy imaging
Improving FRET efficiency measurement in confocal microscopy imaging
复制标题
提高共焦显微镜成像中的 FRET 效率测量
DOI：
10.3788/col20100810.0947
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发表时间：
2010-10
期刊：
影响因子：
--
通讯作者：
Chen, Tongsheng
中科院分区：
文献类型：
--
作者：
Qu, Junle;Yu, Huaina;Chen, Tongsheng
光谱透光率（SBT）取决于共聚焦成像中的荧光强度水平。精密福斯特共振能量转移（FRET）算法根据荧光强度校正SBT比，避免了SBT比的高估或低估。本文提出了一种将精确FRET算法中SBT的计算与E-FRET公式相结合，精确测量单个活细胞中FRET质粒的FRET效率的新方法。我们也用这种方法测量了FRET- bid的FRET效率，发现在健康的A549细胞中，FRET效率约为15%，通过FRET受体光漂白法验证了这一点。
Spectral bleedthrough (SBT) ratio is dependent on the level of fluorescence intensity in confocal imaging. Precision Forster resonance energy transfer (FRET) algorithm corrects SBT ratio according to fluorescence intensity and avoids over- or under-estimation of SBT ratio. In this letter, we propose a new method to accurately measure the FRET efficiency of FRET plasmid in single living cells by combining the calculation of SBT in precision FRET algorithm with E-FRET formulae. We also use this method to measure the FRET efficiency of FRET-Bid, and find that in healthy A549 cells it is about 15%, which is verified by FRET acceptor photobleaching method.
影响因子：
2.8
作者：
R. Stein
通讯作者：
R. Stein