A Membrane-Associated FRET Sensor for Detection of Apoptosis

Herbert Schneckenburger; Michael Wagner; Petra Weber; Thomas Bruns; Heiko Steuer; Brigitte Angres

doi:10.1364/ECBO.2009.7368_0Q

Clinical and Biomedical Spectroscopy
Proceedings of SPIE-OSA Biomedical Optics (Optica Publishing Group, 2009),
paper 7368_0Q
•https://doi.org/10.1364/ECBO.2009.7368_0Q

A Membrane-Associated FRET Sensor for Detection of Apoptosis

Herbert Schneckenburger, Michael Wagner, Petra Weber, Thomas Bruns, Heiko Steuer, and Brigitte Angres

European Conference on Biomedical Optics 2009

Munich Germany
14–18 June 2009
ISBN: 9780819476449

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BIOSPECTROSCOPY AND POINT-OF-CARE DIAGNOSTICS I (BPDI)

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H. Schneckenburger, M. Wagner, P. Weber, T. Bruns, H. Steuer, and B. Angres, "A Membrane-Associated FRET Sensor for Detection of Apoptosis," in Clinical and Biomedical Spectroscopy, I. Georgakoudi, J. Popp, and K. Svanberg, eds., Vol. 7368 of Proceedings of SPIE-OSA Biomedical Optics (Optica Publishing Group, 2009), paper 7368_0Q.

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Abstract

A membrane associated caspase sensor based on Förster Resonance Energy Transfer (FRET) between enhanced cyan fluorescent protein (Mem-ECFP) and yellow fluorescent protein (EYFP) is reported. Upon apoptosis a caspase sensitive amino acid peptide linker (DEVD) between these proteins is cleaved, and pronounced changes of fluorescence spectra and lifetimes are observed. Membrane selective detection of fluorescent proteins in cultivated HeLa cervix carcinoma cells is achieved by total internal reflection fluorescence microscopy (TIRFM) with high sensitivity and resolution.

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