Intensity range based quantitative FRET data analysis to localize protein molecules in live cell nuclei

被引:43
作者
Chen, Y
Periasamy, A
机构
[1] Univ Virginia, WM Keck Ctr Cellular Imaging, Charlottesville, VA 22904 USA
[2] Univ Virginia, Dept Biol, Charlottesville, VA 22904 USA
[3] Univ Virginia, Dept Biomed Engn, Charlottesville, VA 22904 USA
关键词
FRET; spectral bleedthrough (SBT); protein-protein interactions; green fluorescent proteins (GFPs);
D O I
10.1007/s10895-005-0024-1
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Forster (fluorescence) resonance energy transfer (FRET) is an ideal technique to estimate the distance between interacting protein molecules in live specimens using intensity-based microscopy. The spectral overlap of donor and acceptor- essential for FRET-also generates a contamination of the FRET signal. There are a number of algorithms available to remove this spectral bleedthrough (SBT) contamination and in this paper we compare two popular algorithms to estimate the SBT element and to calculate a more precise level of energy transfer efficiency, and with that a more accurate distance estimate.
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页码:95 / 104
页数:10
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