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Keywords:

  • evanescent field;
  • membrane imaging;
  • super-resolution;
  • label-free samples;
  • multilayer Fresnel model;
  • Total Internal Reflection microscopy

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. (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)