Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics.
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.
Shroff
Hari
H
Janelia Farm Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA.
Galbraith
Catherine G
CG
Galbraith
James A
JA
Betzig
Eric
E
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008853:Microscopy / Q000379:methods*; D036103:Nanotechnology; D051419:Paxillin / Q000378:metabolism; D021381:Protein Transport; D013194:Staining and Labeling
D051419:Paxillin