7 May 1997 Time-resolved DNA identification in capillary gel electrophoresis with semiconductor lasers
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Proceedings Volume 2980, Advances in Fluorescence Sensing Technology III; (1997) https://doi.org/10.1117/12.273569
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States
Abstract
We developed a simple, tiny setup with only a few optical devices for fast and sensitive identification of DNA with a short-pulse semiconductor laser operating at 20 MHz. In combination with newly synthesized fluorescent dyes (rhodamine derivatives) which exhibit high fluorescence quantum efficiencies and distinct fluorescence lifetimes at semiconductor laser excitation wavelength a sensitive detection and time-resolved identification of DNA can be achieved. At an excitation wavelength of 635 nm the fluorescence background is greatly reduced. We demonstrate the DNA identification of A- and G-terminated DNA fragments labeled at the 5'-end with the rhodamine derivatives MR 200- 1 and JA 169 during capillary gel electrophoresis. The characteristic time-resolved data are acquired by the time- correlated single-photon counting technique. Time-resolved identification analysis is realized by the maximum likelihood estimator. For prediction of the error rate (misclassification) Cramers equation in combination with a pattern recognition technique is applied. These methods deliver high reliabilities at low classification error rates for low fluorescence light level applications.
© (1997) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Ralph Mueller, Ralph Mueller, Dirk Herten, Dirk Herten, Ulrike Lieberwirth, Ulrike Lieberwirth, Michael Neumann, Michael Neumann, Markus Sauer, Markus Sauer, Andreas Schulz, Andreas Schulz, Stefan Siebert, Stefan Siebert, Karl-Heinz Drexhage, Karl-Heinz Drexhage, Juergen M. Wolfrum, Juergen M. Wolfrum, } "Time-resolved DNA identification in capillary gel electrophoresis with semiconductor lasers", Proc. SPIE 2980, Advances in Fluorescence Sensing Technology III, (7 May 1997); doi: 10.1117/12.273569; https://doi.org/10.1117/12.273569
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