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Journal of Biomedical Optics

Two-photon interactions at single fluorescent molecule level
Author(s): Fabio Cannone; Giuseppe Chirico; Alberto Diaspro
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Paper Abstract

Single-molecule spectroscopy and single-molecule detection are emerging areas that have many applications when combined with scanning, imaging, and spectroscopy techniques. We have combined a commercial confocal scanning head, to a Ti:sapphire laser and to an inverted microscope, for the detection of single molecule fluorescence of varies dyes by two-photon excitation. We collected spot images of fluorescent molecules that have been deposited on a substrate considering both blinking and photobleaching behavior of fluorescent spots. Here, we report data related to two-photon interactions that occur with the following fluorescent molecules: Indo-1, Rhodamine 6G, Fluorescein, and Pyrene. The choice of these specific dyes is based upon their wide use in biological and medical applications together with the varying complexity of their chemical structure that increases from Pyrene to Indo-1. Moreover, we report some data about single molecule studies related to denaturation of an enhanced green fluorescent protein, GFPmut2, under one photon excitation regime, that show a very similar trend to that observed for the already mentioned fluorescent molecules.

Paper Details

Date Published: 1 July 2003
PDF: 5 pages
J. Biomed. Opt. 8(3) doi: 10.1117/1.1582180
Published in: Journal of Biomedical Optics Volume 8, Issue 3
Show Author Affiliations
Fabio Cannone, Univ. degli Studi di Milano-Bicocca (Italy)
Giuseppe Chirico, Univ. degli Studi di Milano-Bicocca (Italy)
Alberto Diaspro, Univ. degli Studi di Genova (Italy)

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