Two-photon interactions at single fluorescent molecule level

被引:7
作者
Cannone, F [1 ]
Chirico, G
Diaspro, A
机构
[1] Natl Inst Phys Matter, Lab Adv Microscopy Bioimaging & Spect, Genoa, Italy
[2] Univ Milano Bicocca, Dept Phys, I-20126 Milan, Italy
[3] Univ Genoa, Dept Phys, I-16146 Genoa, Italy
关键词
single-molecule detection; fluorescence detection; two-photon excitation; thermal bleaching; two-photon interactions; fluorescent molecules;
D O I
10.1117/1.1582180
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 for the detection of single molecule and to an inverted microscope, 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. (C) 2003 Society of Photo-Optical Instrumentation Engineers.
引用
收藏
页码:391 / 395
页数:5
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