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

Analytical Chemistry
Qiang Li, Stefan Seeger

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.

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Citations

Oct 5, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Jian ZhangJoseph R Lakowicz
Jun 8, 2007·Analytical Biochemistry·Qiang Li, Stefan Seeger
Jul 13, 2012·Biosensors & Bioelectronics·Kyoko FukazawaKazuhiko Ishihara
Oct 6, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Gamze Kavran BelinStefan Seeger
Jun 11, 2019·Physical Chemistry Chemical Physics : PCCP·Ismail A AhmedFeng Gai
May 23, 2008·Analytical Chemistry·Mark LowryIsiah M Warner
Oct 15, 2011·The Journal of Physical Chemistry. B·Qiang Li, Stefan Seeger
Feb 23, 2020·The Journal of Physical Chemistry Letters·Aleksandr Barulin, Jérôme Wenger
Feb 8, 2011·Advances in Colloid and Interface Science·Michael RabeStefan Seeger

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