Two-photon interactions at single fluorescent molecule level

Fabio Cannone; Giuseppe Chirico; Alberto Diaspro

doi:10.1117/1.1582180

1 July 2003 Two-photon interactions at single fluorescent molecule level

Fabio Cannone, Giuseppe Chirico, Alberto Diaspro

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Journal of Biomedical Optics, Vol. 8, Issue 3, (July 2003). https://doi.org/10.1117/1.1582180

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.

©(2003) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Fabio Cannone, Giuseppe Chirico, and Alberto Diaspro "Two-photon interactions at single fluorescent molecule level," Journal of Biomedical Optics 8(3), (1 July 2003). https://doi.org/10.1117/1.1582180

Published: 1 July 2003

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KEYWORDS

Luminescence

Molecules

Glasses

Molecular interactions

Microscopes

Rhodamine

Confocal microscopy

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