使用深紫外荧光寿命显微镜对单个蛋白质分子进行无标记检测。

Label-free detection of single protein molecules using deep UV fluorescence lifetime microscopy.

作者信息

Li Qiang, Seeger Stefan

机构信息

Physikalisch-Chemisches Institut der Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.

出版信息

Anal Chem. 2006 Apr 15;78(8):2732-7. doi: 10.1021/ac052166u.

DOI:10.1021/ac052166u

PMID:16615786

Abstract

We present the detection of single beta-galactosidase molecules from Escherichia coli (Ecbeta Gal) using deep UV laser-based fluorescence lifetime microscopy. The native fluorescence from intrinsic tryptophan emission has been observed after one-photon excitation at 266 nm. Applying the time-resolved single-photon counting method, we investigated the fluorescence lifetime distribution and the bursts of autofluorescence photons from tryptophan residues in Ecbeta Gal protein as well as fluorescence correlation spectroscopy of Ecbeta Gal. The results demonstrate that deep UV laser-based fluorescence lifetime microscopy is useful for identification of biological macromolecules at the single-molecule level using intrinsic fluorescence.

摘要

我们展示了使用基于深紫外激光的荧光寿命显微镜对大肠杆菌β-半乳糖苷酶（Ecbeta Gal）单个分子的检测。在266nm的单光子激发后，观察到了来自内在色氨酸发射的天然荧光。应用时间分辨单光子计数方法，我们研究了Ecbeta Gal蛋白中色氨酸残基的荧光寿命分布和自发荧光光子的猝发，以及Ecbeta Gal的荧光相关光谱。结果表明，基于深紫外激光的荧光寿命显微镜对于使用内在荧光在单分子水平鉴定生物大分子是有用的。