We have characterized a commercial confocal scanning head for the detection of single molecule fluorescence by two-photon excitation. We have verified that the distribution of the fluorescence emitted by dyes and labeled proteins on glass substrates is discrete with quanta proportional to a common reference signal. We describe and test a simple and quantitative tool to discriminate between single molecules and molecular aggregates on single snapshots based on the analysis of the intensity distribution. We have verified the square dependence of the fluorescence intensity vs. the excitation power, suggesting that no appreciable saturation and fast photo-damage of the chromophores takes place at the excitation power employed here. (C) 2001 Wiley-Liss. Inc.

Chirico, G., Cannone, F., Beretta, S., Baldini, G., Diaspro, A. (2001). Single molecule studies by means of the two-photon fluorescence distribution. MICROSCOPY RESEARCH AND TECHNIQUE, 55(5), 359-364 [10.1002/jemt.10003].

Single molecule studies by means of the two-photon fluorescence distribution

CHIRICO, GIUSEPPE;BALDINI, GIANCARLO;
2001

Abstract

We have characterized a commercial confocal scanning head for the detection of single molecule fluorescence by two-photon excitation. We have verified that the distribution of the fluorescence emitted by dyes and labeled proteins on glass substrates is discrete with quanta proportional to a common reference signal. We describe and test a simple and quantitative tool to discriminate between single molecules and molecular aggregates on single snapshots based on the analysis of the intensity distribution. We have verified the square dependence of the fluorescence intensity vs. the excitation power, suggesting that no appreciable saturation and fast photo-damage of the chromophores takes place at the excitation power employed here. (C) 2001 Wiley-Liss. Inc.
Articolo in rivista - Articolo scientifico
single molecules; two-photon excitation; fluorescence microscopy; intensity distributions; Betalactoglobulin; rhodamine 6G; fluorescein
English
2001
55
5
359
364
none
Chirico, G., Cannone, F., Beretta, S., Baldini, G., Diaspro, A. (2001). Single molecule studies by means of the two-photon fluorescence distribution. MICROSCOPY RESEARCH AND TECHNIQUE, 55(5), 359-364 [10.1002/jemt.10003].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/17182
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