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Interference Reflection Microscopy

  1. Igor Weber

Published Online: 17 JAN 2011

DOI: 10.1002/9780470015902.a0002636.pub2

eLS

eLS

How to Cite

Weber, I. 2011. Interference Reflection Microscopy. eLS. .

Author Information

  1. Ruder Bošković Institute, Division of Molecular Biology, Zagreb, Croatia

Publication History

  1. Published Online: 17 JAN 2011

Abstract

Interference reflection microscopy (IRM) utilises interference of light reflected from closely apposed surfaces to provide an image containing information about the separation of those surfaces. In cell biology, IRM is used to image structures at the base of adherent cells and to measure cell–substratum distances, as well as to investigate mechanisms of cell–substratum adhesion. IRM is also used to study topology and dynamics of biomimetic systems such as vesicles, supported membranes and other multilayered structures. Basic IRM optical configuration is relatively easy to set up, and image analysis can provide information about interfacial distances with nanometer precision and millisecond time resolution. IRM can be readily combined with other microscopic techniques, and with force transducing devices such as optical tweezers, micropipettes and microcantilevers. New advancements in the field include dual-wavelength IRM and fluctuation contrast IRM.

Key Concepts:

  • Interference reflection microscopy measures the distance between close surfaces.

  • Cell adhesion areas such as focal contacts can be mapped by IRM.

  • IRM provides vertical resolution in the nanometer range.

  • Dual-wavelength IRM removes ambiguity in measurements of vertical distances up to 800 nm.

  • IRM can determine amplitudes of local membrane fluctuations.

Keywords:

  • IRM;
  • RICM;
  • reflection interference contrast;
  • cell adhesion;
  • cell–substratum contact;
  • microinterferometry;
  • interfacial distance measurements;
  • membrane fluctuations;
  • focal contacts