Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET)

Biochem Soc Trans. 2003 Oct;31(Pt 5):1020-7. doi: 10.1042/bst0311020.

Abstract

We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow cytometry of cells. These methods permit the assessment of rotational motion, association and proximity of cellular proteins in vivo. They are particularly applicable to probes generated by fusions of visible fluorescence proteins, as exemplified by studies of the erbB receptor tyrosine kinases involved in growth-factor-mediated signal transduction.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Review

MeSH terms

  • Animals
  • Anisotropy*
  • CHO Cells
  • Cell Line
  • Cricetinae
  • Dose-Response Relationship, Drug
  • ErbB Receptors / chemistry
  • Flow Cytometry
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods*
  • Models, Statistical
  • Mutation

Substances

  • Luminescent Proteins
  • Green Fluorescent Proteins
  • ErbB Receptors