Imaging of protein cluster sizes by means of confocal time-gated fluorescence anisotropy microscopy

被引：40

作者：

Bader, Arjen N. ^{[1
]}

Hofman, Erik G. ^{[1
]}

Henegouwen, Paul M. P. van Bergen en ^{[1
]}

Gerritsen, Hans C. ^{[1
]}

机构：

[1] Univ Utrecht, NL-3508 TA Utrecht, Netherlands

来源：

OPTICS EXPRESS | 2007年 / 15卷 / 11期

关键词：

D O I：

10.1364/OE.15.006934

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

A time-resolved fluorescence anisotropy imaging method for studying nanoscale clustering of proteins or lipids was developed and evaluated. It is based on FRET between the identical fluorophores (homo-FRET), which results in a rapid depolarization of the fluorescence. The method employs the time-resolved fluorescence anisotropy decays recorded in a confocal microscope equipped with pulsed excitation and time-gated detection. From the decay the limiting anisotropy r(inf) was derived, which is a direct measure for the number of fluorophores per cluster. The method was evaluated by imaging GPI-GFP, a lipid raft marker. Small clusters were observed in the plasma membrane while the cytoplasm and the Golgi contained predominantly monomers. (C) 2007 Optical Society of America.

引用

页码：6934 / 6945

页数：12

共 25 条

[21] Time-resolved fluorescence anisotropy imaging applied to live cells [J].