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13 February 2012 Quantifying the influence of yellow fluorescent protein photoconversion on acceptor photobleaching-based fluorescence resonance energy transfer measurements
Arne Seitz, Stefan Terjung, Timo Zimmermann, Rainer Pepperkok
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Journal of Biomedical Optics, Vol. 17, Issue 1, 011010 (February 2012). https://doi.org/10.1117/1.JBO.17.1.011010
Abstract
Fluorescence resonance energy transfer (FRET) efficiency measurements based on acceptor photobleaching of yellow fluorescent protein (YFP) are affected by the fact that bleaching of YFP produces a fluorescent species that is detectable in cyan fluorescent protein (CFP) image channels. The presented quantitative measurement of this conversion makes it possible to correct the obtained FRET signal to increase the accuracy of intensity based CFP/YFP FRET measurements. The described method can additionally be used to compare samples with very different fluorescence levels.
© 2012 Society of Photo-Optical Instrumentation Engineers (SPIE) 0091-3286/2012/$25.00 © 2012 SPIE
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Arne Seitz, Stefan Terjung, Timo Zimmermann, and Rainer Pepperkok "Quantifying the influence of yellow fluorescent protein photoconversion on acceptor photobleaching-based fluorescence resonance energy transfer measurements," Journal of Biomedical Optics 17(1), 011010 (13 February 2012). https://doi.org/10.1117/1.JBO.17.1.011010
Published: 13 February 2012
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Cited by 9 scholarly publications.
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KEYWORDS
Fluorescence resonance energy transfer
Fluorescent proteins
Confocal microscopy
Luminescence
Microscopes
Calibration
Microscopy
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