Identification of single molecules in aqueous solution by time-resolved fluorescence anisotropy

被引：142

作者：

Schaffer, J ^{[1
]}

Volkmer, A ^{[1
]}

Eggeling, C ^{[1
]}

Subramaniam, V ^{[1
]}

Striker, G ^{[1
]}

Seidel, CAM ^{[1
]}

机构：

[1] Max Planck Inst Biophys Chem, D-37077 Gottingen, Germany

来源：

JOURNAL OF PHYSICAL CHEMISTRY A | 1999年 / 103卷 / 03期

关键词：

D O I：

10.1021/jp9833597

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

Using a confocal epi-illuminated microscope with a polarizing beam splitter and dual-channel detection of single-molecule fluorescence induced by pulsed laser excitation, a new application of the three-dimensional, real-time spectroscopic technique BIFL (burst integrated fluorescence lifetime) is introduced. BIFL allows simultaneous registration of fluorescence intensity, lifetime, and anisotropy. It is shown to be well-suited to identify the freely diffusing fluorescent molecule Rhodamine 123 and the Enhanced Yellow Fluorescent Protein via their characteristic fluorescence anisotropy using a time-resolved analysis. Furthermore, data analysis is discussed and rotational correlation times of single molecules are determined. Applications for multidimensional single-molecule identification are outlined.

引用

页码：331 / 336

页数：6

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