Diode-laser-based time-resolved detection and identification of individual mononucleotide molecules in aqueous solution

Markus Sauer; Christoph Zander; Ralph Mueller; Florian Goebel; Andreas Schulz; Stefan Siebert; Karl-Heinz Drexhage; Juergen M. Wolfrum

doi:10.1117/12.274338

22 May 1997 Diode-laser-based time-resolved detection and identification of individual mononucleotide molecules in aqueous solution

Markus Sauer, Christoph Zander, Ralph Mueller, Florian Goebel, Andreas Schulz, Stefan Siebert, Karl-Heinz Drexhage, Juergen M. Wolfrum

Author Affiliations +

Proceedings Volume 2985, Ultrasensitive Biochemical Diagnostics II; (1997) https://doi.org/10.1117/12.274338
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

We applied a short-pulse diode laser emitting at 637 nm with a repetition rate of 30 MHz in combination with a confocal microscope to study bursts of fluorescence photons from individual labeled mononucleotide molecules in water. A newly synthesized oxazine dye and the commercially available carbocyanine dye Cy5 were used as fluorescent labels. Multichannel scalar traces, the fluorescence autocorrelation function and fluorescence decay times determined by time- correlated single-photon counting have been measured simultaneously. The time-resolved signals of the two mononucleotides were analyzed and identified by a maximum likelihood estimator. The results showed out that 60 detected photons per transit of a single molecule are sufficient to distinguish two labeled mononucleotides in water with a misclassification of less than 10 percent via their characteristic fluorescence lifetimes of 1.07 +/- 0.27 ns and 1.89 +/- 0.34 ns.

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Markus Sauer, Christoph Zander, Ralph Mueller, Florian Goebel, Andreas Schulz, Stefan Siebert, Karl-Heinz Drexhage, and Juergen M. Wolfrum "Diode-laser-based time-resolved detection and identification of individual mononucleotide molecules in aqueous solution", Proc. SPIE 2985, Ultrasensitive Biochemical Diagnostics II, (22 May 1997); https://doi.org/10.1117/12.274338

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