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Journal of Biomedical Optics

Robust single-molecule approach for counting autofluorescent proteins
Author(s): Laurent Cognet; Catherine Tardin; Marie-Laure Martin Negrier; Christelle Breillat; Francoise Coussen; Daniel Choquet; Brahim Lounis
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Paper Abstract

Using single-molecule microscopy, we present a method to quantify the number of single autofluorescent proteins when they cannot be optically resolved. This method relies on the measurement of the total intensity emitted by each aggregate until it photobleaches. This strategy overcomes the inherent problem of blinking of green fluorescent proteins. In the case of small protein aggregates, our method permits us to describe the mean composition with a precision of one protein. For aggregates containing a large number of proteins, it gives access to the average number of proteins gathered and a signature of the inhomogeneity of the aggregates' population. We applied this methodology to the quantification of small purified citrine multimers.

Paper Details

Date Published: 1 May 2008
PDF: 5 pages
J. Biomed. Opt. 13(3) 031216 doi: 10.1117/1.2940600
Published in: Journal of Biomedical Optics Volume 13, Issue 3
Show Author Affiliations
Laurent Cognet, Univ. Bordeaux I (France)
Catherine Tardin, Univ. Bordeaux I (France)
Marie-Laure Martin Negrier, Univ. Bordeaux I (France)
Christelle Breillat, Univ. Bordeaux I (France)
Francoise Coussen, Univ. Victor Segalen Bordeaux 2 (France)
Daniel Choquet, Univ. Victor Segalen Bordeaux 2 (France)
Brahim Lounis, Univ. Bordeaux I (France)

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