Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics

被引：528

作者：

Shroff, Hari ^{[1
]}

Galbraith, Catherine G. ^{[2
]}

Galbraith, James A. ^{[3
]}

Betzig, Eric ^{[1
]}

机构：

[1] Howard Hughes Med Inst, Ashburn, VA 20147 USA

[2] Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD 20892 USA

[3] Natl Inst Neurol Disorders & Stroke, NIH, Bethesda, MD 20892 USA

来源：

NATURE METHODS | 2008年 / 5卷 / 05期

关键词：

D O I：

10.1038/NMETH.1202

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We demonstrate live-cell super-resolution imaging using photoactivated localization microscopy (PALM). The use of photon-tolerant cell lines in combination with the high resolution and molecular sensitivity of PALM permitted us to investigate the nanoscale dynamics within individual adhesion complexes (ACs) in living cells under physiological conditions for as long as 25 min, with half of the time spent collecting the PALM images at spatial resolutions down to similar to 60 nm and frame rates as short as 25 s. We visualized the formation of ACs and measured the fractional gain and loss of individual paxillin molecules as each AC evolved. By allowing observation of a wide variety of nanoscale dynamics, live-cell PALM provides insights into molecular assembly during the initiation, maturation and dissolution of cellular processes.

引用

收藏

页码：417 / 423

页数：7

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