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Nat Methods. 2008 May;5(5):417-23. doi: 10.1038/nmeth.1202. Epub 2008 Apr 13.

Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics.

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1
Janelia Farm Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA.

Abstract

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 approximately 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.

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PMID:
18408726
PMCID:
PMC5225950
DOI:
10.1038/nmeth.1202
[Indexed for MEDLINE]
Free PMC Article
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