Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics

Hari Shroff, Catherine G. Galbraith, James A. Galbraith, Eric Betzig

Research output: Contribution to journalArticle

589 Scopus citations

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 ∼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.

Original languageEnglish (US)
Pages (from-to)417-423
Number of pages7
JournalNature Methods
Volume5
Issue number5
DOIs
StatePublished - May 2008

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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