Dynamic superresolution imaging of endogenous proteins on living cells at ultra-high density.
Biophysical Journal. 2010-08-01; 99(4): 1303-1310
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1. Biophys J. 2010 Aug 9;99(4):1303-10. doi: 10.1016/j.bpj.2010.06.005.
Dynamic superresolution imaging of endogenous proteins on living cells at
Giannone G(1), Hosy E, Levet F, Constals A, Schulze K, Sobolevsky AI, Rosconi MP,
Gouaux E, Tampé R, Choquet D, Cognet L.
(1)Centre National de la Recherche Scientifique UMR 5091, Cellular Physiology of
the Synapse, Bordeaux, France.
Nat Methods. 2010 Oct;7(10):784.
Versatile superresolution imaging methods, able to give dynamic information of
endogenous molecules at high density, are still lacking in biological science.
Here, superresolved images and diffusion maps of membrane proteins are obtained
on living cells. The method consists of recording thousands of single-molecule
trajectories that appear sequentially on a cell surface upon continuously
labeling molecules of interest. It allows studying any molecules that can be
labeled with fluorescent ligands including endogenous membrane proteins on living
cells. This approach, named universal PAINT (uPAINT), generalizes the previously
developed point-accumulation-for-imaging-in-nanoscale-topography (PAINT) method
for dynamic imaging of arbitrary membrane biomolecules. We show here that the
unprecedented large statistics obtained by uPAINT on single cells reveal local
diffusion properties of specific proteins, either in distinct membrane
compartments of adherent cells or in neuronal synapses.
2010 Biophysical Society. Published by Elsevier Inc. All rights reserved.
PMID: 20713016 [Indexed for MEDLINE]