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Label‐free evanescent microscopy for membrane nano‐tomography in living cells
Author(s) -
Bon Pierre,
Barroca Thomas,
LévèqueFort Sandrine,
Fort Emmanuel
Publication year - 2014
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201300095
Subject(s) - membrane , microscopy , microscope , tomography , live cell imaging , tracking (education) , nano , living cell , nanotechnology , materials science , optics , chemistry , biophysics , biological system , cell , physics , biology , biochemistry , psychology , pedagogy , composite material
We show that through‐the‐objective evanescent microscopy (epi‐EM) is a powerful technique to image membranes in living cells. Readily implementable on a standard inverted microscope, this technique enables full‐field and real‐time tracking of membrane processes without labeling and thus signal fading. In addition, we demonstrate that the membrane/interface distance can be retrieved with 10 nm precision using a multilayer Fresnel model. We apply this nano‐axial tomography of living cell membranes to retrieve quantitative information on membrane invagination dynamics. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)