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FRET: A Spectral Ruler for Interacting Molecules Involved in Apoptosis

Published online by Cambridge University Press:  02 July 2020

Victoria E. Centonze
Affiliation:
Department of Cellular and Structural Biology, University of Texas HealthScience Center at San Antonio, San Antonio, TX78229-3900.
Xiao H. Liang
Affiliation:
Department of Medicine, Columbia University College of Physicians &Surgeons, New York, NY10032.
Elizabeth A. Casanova
Affiliation:
Department of Cellular and Structural Biology, University of Texas HealthScience Center at San Antonio, San Antonio, TX78229-3900.
Beth Levine
Affiliation:
Department of Medicine, Columbia University College of Physicians &Surgeons, New York, NY10032.
Brian Herman
Affiliation:
Department of Cellular and Structural Biology, University of Texas HealthScience Center at San Antonio, San Antonio, TX78229-3900.
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Extract

Fluorescence Resonance Energy Transfer (FRET) is a process by which a fluorophore (the donor) in the excited state transfers its energy to a neighboring fluorophore (the acceptor) non-radiatively through dipole-dipole interactions. Since the efficiency of energy transfer varies as the inverse of the sixth power of the distance separating the donor and acceptor chromophores, for FRET to occur the distance between the two molecules cannot exceed 10 to 100 angstroms (1 to l0nm). The combination of FRET and optical microscopy allows examination and quantitation of dynamic molecular interactions between cellular constituents at resolutions beyond the Abbe diffraction limit of light microscopy. Through the microscope one may detect FRET by an overall decrease in fluorescence emission of the donor with a concomitant increase in fluorescence emission of the acceptor.

Type
Recent Advances in Light Microscopy
Copyright
Copyright © Microscopy Society of America

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