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

被引：25

作者：

Chirico, G

Cannone, F

Beretta, S

Baldini, G

Diaspro, A

机构：

[1] Ist Nazl Fis Mat, UdR Milano Bicocca, I-20126 Milan, Italy

[2] UdR Genova, I-16146 Genoa, Italy

来源：

MICROSCOPY RESEARCH AND TECHNIQUE | 2001年 / 55卷 / 05期

关键词：

single molecules; two-photon excitation; fluorescence microscopy; intensity distributions; Betalactoglobulin; rhodamine; 6G; fluorescein;

D O I：

10.1002/jemt.10003

中图分类号：

R602 [外科病理学、解剖学]; R32 [人体形态学];

学科分类号：

100101 ;

摘要：

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.

引用

页码：359 / 364

页数：6

共 15 条

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Ambrose WP, 1997, CHEM PHYS LETT, V269, P365, DOI 10.1016/S0009-2614(97)00266-2

[2] Direct imaging single molecule diffusion in a solid polymer host [J].