Basics of Fluorescence and FRET

Research output: Chapter in Book/Report/Conference proceedingChapter

22 Citations (Scopus)

Abstract

This chapter discusses the most important characteristics of fluorescence that plays a fundamental role in understanding the basics and the applications of Förster (fluorescence) resonance (radiationless) energy transfer (FRET). FRET is the transfer of electronic excitation energy between isolated donor D and acceptor A of suitable spectroscopic properties. The donor molecules, typically, emit at shorter wavelengths, which overlap with the absorption spectrum of the acceptor. This energy transfer occurs without the appearance of the photon and is the result of long-range interactions between the D and A dipoles. The most important factors affecting FRET are the overlap integral, the quantum yield of the donor in the absence of the acceptor, and the orientation factor. The quantitative analysis of steady-state and time-resolved FRET measurements provides information on global structures and conformational dynamics, and reveals thermodynamic parameters for conformational transition. This information is essential for the understanding of biological functions of proteins, DNA/RNA, and other biological assemblies that are frequently mediated by transitions between alternative conformations.

Original languageEnglish
Title of host publicationMolecular Imaging
PublisherElsevier Inc.
Pages21-56
Number of pages36
ISBN (Print)9780195177206
DOIs
StatePublished - 1 Dec 2005

Fingerprint

Fluorescence Resonance Energy Transfer
Excitation energy
Energy Transfer
Quantum yield
RNA
Photons
Thermodynamics
Energy transfer
Conformations
Absorption spectra
DNA
Fluorescence
energy transfer
Proteins
Wavelength
fluorescence
Molecules
resonance fluorescence
Chemical analysis
assemblies

Cite this

Gryczynski, Z., Gryczynski, I., & Lakowicz, J. R. (2005). Basics of Fluorescence and FRET. In Molecular Imaging (pp. 21-56). Elsevier Inc.. https://doi.org/10.1016/B978-019517720-6.50011-0
Gryczynski, Zygmunt ; Gryczynski, Ignacy ; Lakowicz, Joseph R. / Basics of Fluorescence and FRET. Molecular Imaging. Elsevier Inc., 2005. pp. 21-56
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Gryczynski, Z, Gryczynski, I & Lakowicz, JR 2005, Basics of Fluorescence and FRET. in Molecular Imaging. Elsevier Inc., pp. 21-56. https://doi.org/10.1016/B978-019517720-6.50011-0

Basics of Fluorescence and FRET. / Gryczynski, Zygmunt; Gryczynski, Ignacy; Lakowicz, Joseph R.

Molecular Imaging. Elsevier Inc., 2005. p. 21-56.

Research output: Chapter in Book/Report/Conference proceedingChapter

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AB - This chapter discusses the most important characteristics of fluorescence that plays a fundamental role in understanding the basics and the applications of Förster (fluorescence) resonance (radiationless) energy transfer (FRET). FRET is the transfer of electronic excitation energy between isolated donor D and acceptor A of suitable spectroscopic properties. The donor molecules, typically, emit at shorter wavelengths, which overlap with the absorption spectrum of the acceptor. This energy transfer occurs without the appearance of the photon and is the result of long-range interactions between the D and A dipoles. The most important factors affecting FRET are the overlap integral, the quantum yield of the donor in the absence of the acceptor, and the orientation factor. The quantitative analysis of steady-state and time-resolved FRET measurements provides information on global structures and conformational dynamics, and reveals thermodynamic parameters for conformational transition. This information is essential for the understanding of biological functions of proteins, DNA/RNA, and other biological assemblies that are frequently mediated by transitions between alternative conformations.

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