Microscopic Analysis of Fluorescence Resonance Energy Transfer (FRET)

  • Brian Herman
  • R. Venkata Krishnan
  • Victoria E. Centonze
Part of the Methods in Molecular Biology book series (MIMB, volume 261)


We describe here detailed practical procedures for implementing various approaches of fluorescence resonance energy transfer (FRET) in microscope-based measurements. A comprehensive theoretical formalism is developed and the different experimental procedures are outlined. A step-by-step protocol is provided for preparing the specimens for FRET measurements, data acquisition procedures, analysis, and quantification. Particular emphasis is given to exemplify the FRET applications in the study of protein-protein interactions.

Key Words

Microscopy FRET FLIM protein interactions 


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Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Brian Herman
    • 1
  • R. Venkata Krishnan
    • 1
  • Victoria E. Centonze
    • 1
  1. 1.Department of Cellular & Structural BiologyUniversity of Texas Health Science CenterSan Antonio

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