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  Photochromic Fluorescence Resonance Energy Transfer (pcFRET): formalism, implementation, and perspectives

Jares-Erijman, E. A., Giordano, L., Spagnuolo, C., Kawior, J., Vermeij, R. J., & Jovin, T. M. (2004). Photochromic Fluorescence Resonance Energy Transfer (pcFRET): formalism, implementation, and perspectives. In Periasamy, A., S. P. T., & Society of Photo-Optical Instrumentation Engineers (Eds.), Multiphoton microscopy in the biomedical sciences IV; Conference, San Jose, Calif., 25 - 27 January 2004 (pp. 13-26). Bellingham, WA: SPIE Publ.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-EEAF-D Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-E2FE-D
Genre: Book Chapter

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 Creators:
Jares-Erijman, E. A.1, Author              
Giordano, L., Author
Spagnuolo, C.1, Author              
Kawior, J., Author
Vermeij, R. J.1, Author              
Jovin, T. M.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: FRET, adFRET , dithienylethene, Quantum Dot, chemical equilibria, relaxation kinetics, pcRelKin
 Abstract: Photochromic FRET (pcFRET), a member of the family of acceptor epletion FRET techniques (adFRET), embodies a general conceptual and experimental scheme based on a coupled system of a fluorescent donor and a photchromic acceptor. The procedure involves the reversible and cyclic spectroscopic depletion of the acceptor, and was initially conceived for the determination of FRET efficiency on a continuos, pixel-by-pixel basis in the microscopy of living cells. However, the modulation of donor fluorescence in pcFRET has implications for a wide range of applications. We present the formalism for quantitative interpretations of photostationary and kinetic data, from which the relevant kinetic rate constants and quantum uields for the cyclization and cycloreversion reactions of the photochromic acceptor can be derived. The scheme was applied to a model system consisting of a fluorescent donor (Lucifer Yellow) covalently bound to a diheteroarylethene acceptor. In a Perspectives section, we discuss photochromic probes, instrumentation issues, and the potential of pcFRET for analyizing chemical equilibria and kinetics, in the latter case with a new technique we have denoted Photochromic Relaxation Kinetics (pcRelKin).

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Language(s): eng - English
 Dates: 2004-06-302004
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 149376
 Degree: -

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Title: Multiphoton microscopy in the biomedical sciences IV; Conference, San Jose, Calif., 25 - 27 January 2004
Source Genre: Book
 Creator(s):
Periasamy, Editor
A., Editor
T., So. P., Editor
Society of Photo-Optical Instrumentation Engineers, Editor  
Affiliations:
-
Publ. Info: Bellingham, WA : SPIE Publ.
Pages: - Volume / Issue: - Sequence Number: - Start / End Page: 13 - 26 Identifier: -

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Title: SPIE Proceedings; Progress in biomedical optics and imaging
Source Genre: Series
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Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 5323; 5, 12 Sequence Number: - Start / End Page: - Identifier: -