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Journal of Biomedical Optics • Open Access

Förster resonance energy transfer-based total internal reflection fluorescence reader for apoptosis
Author(s): Thomas Bruns; Brigitte Angres; Heiko Steuer; Petra Weber; Michael Wagner; Herbert Schneckenburger

Paper Abstract

A fluorescence reader for the detection of Förster resonance energy transfer (FRET) on surfaces of living cells is described. The method is based on multiple total internal reflections (TIR) of an incident laser beam within a glass slide, such that individual samples on top of the glass slide are illuminated simultaneously by an evanescent electromagnetic field. Enhanced cyan fluorescent protein (ECFP) anchored to the inner leaflet of the plasma membrane is optically excited and transfers its excitation energy via the peptide linker Asp-Glu-Val-Asp (DEVD) to an enhanced yellow fluorescent protein. Upon apoptosis, DEVD is cleaved, and energy transfer is disrupted, as proven by an increase of fluorescence intensity as well as of fluorescence lifetime of the donor ECFP. Due to selective excitation of membrane-associated fluorophores, intracellular fluorescence and background luminescence from the surrounding medium are eliminated. Therefore, this test system appears to be a sensitive device for the detection of apoptosis and more generally for drug screening or in vitro diagnosis on a nanometer scale.

Paper Details

Date Published: 1 March 2009
PDF: 5 pages
J. Biomed. Opt. 14(2) 021003 doi: 10.1117/1.3055622
Published in: Journal of Biomedical Optics Volume 14, Issue 2
Show Author Affiliations
Thomas Bruns, Hochschule Aalen (Germany)
Brigitte Angres, Naturwissenschaftliches und Medizinisches Institut an der Univ. Tübingen (Germany)
Heiko Steuer, Naturwissenschaftliches und Medizinisches Institut an der Univ. Tübingen (Germany)
Petra Weber, Hochschule Aalen (Germany)
Michael Wagner, Hochschule Aalen (Germany)
Herbert Schneckenburger, Hochschule Aalen (Germany)

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