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

来源：

JOURNAL OF FLUORESCENCE | 2006年 / 16卷 / 01期

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.

引用

页码：95 / 104

页数：10

共 32 条

[1] BASOLATERAL TO APICAL TRANSCYTOSIS IN POLARIZED CELLS IS INDIRECT AND INVOLVES BFA AND TRIMERIC-G PROTEIN SENSITIVE PASSAGE THROUGH THE APICAL ENDOSOME [J].