Label-free evanescent microscopy for membrane nano-tomography in living cells

Authors

  • Pierre Bon,

    Corresponding author
    1. Institut Langevin, ESPCI ParisTech, CNRS UMR 7587, 1 rue Jussieu, 75231 Paris cedex 05, France
    2. Institut des Sciences Moléculaires d'Orsay ISMO, Univ. Paris-Sud, CNRS UMR 8214, 91405 Orsay cedex, France
    • Institut Langevin, ESPCI ParisTech, CNRS UMR 7587, 1 rue Jussieu, 75231 Paris cedex 05, France

    Search for more papers by this author
  • Thomas Barroca,

    1. Institut Langevin, ESPCI ParisTech, CNRS UMR 7587, 1 rue Jussieu, 75231 Paris cedex 05, France
    Search for more papers by this author
  • Sandrine Lévèque-Fort,

    1. Institut des Sciences Moléculaires d'Orsay ISMO, Univ. Paris-Sud, CNRS UMR 8214, 91405 Orsay cedex, France
    Search for more papers by this author
  • Emmanuel Fort

    1. Institut Langevin, ESPCI ParisTech, CNRS UMR 7587, 1 rue Jussieu, 75231 Paris cedex 05, France
    Search for more papers by this author

Abstract

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)

Ancillary