Abstract

Research on the time-resolved fluorescence of fluorescein isothiocyanate, NBD, tetramethylrhodamine isothiocyanate, and Texas Red—the dyes used for fluorescence-based DNA sequencing—is described. Mean fluorescence lifetimes in both aqueous buffer solution and 5.3%T, 4.8%C polyacrylamide gel were determined as a function of excitation wavelengths at 337, 470, and 550 nm and were found to be 3.5, 1.1, 2.5, and 4.3 ns; the detection limits are 10, 200, 200, and 200 amol for FITC, NBD, TEMR, and T. Red, respectively. Comparisons of fluorescence parameters between the conjugated dyes and the free dyes are also reported. Results on the optimization of the excitation source wavelengths to improve sensitivity and reduce background scattering in polyacrylamide gel are also reported. Time-resolved fluorescence was successfully applied to resolve spectral overlapping of emissions in both solution and in polyacrylamide gel.

How to find the sensitivity limit for DNA sequencing based on laser-induced fluorescence

Malte Köllner
Appl. Opt. 32(6) 806-820 (1993)

Fluorescence enhancement of hemicyanines bound to DNA or DNA-complex and their application to dye laser

Yuki Suzuki and Yutaka Kawabe
Opt. Mater. Express 7(6) 2062-2068 (2017)

Exciplex liquid-phase thermometer using time-resolved laser-induced fluorescence

Christian Parigger, David H. Plemmons, Ron J. Litchford, and San-Mou Jeng
Opt. Lett. 23(1) 76-78 (1998)

Radiative lifetime measurements in neutral lanthanum using time-resolved laser-induced fluorescence spectroscopy in supersonic free-jet

Suresh Yarlagadda, Sheo Mukund, and S. G. Nakhate
J. Opt. Soc. Am. B 28(8) 1928-1933 (2011)

Time-resolved resonance fluorescence spectroscopy for study of chemical reactions in laser-induced plasmas

Lei Liu, Leimin Deng, Lisha Fan, Xi Huang, Yao Lu, Xiaokang Shen, Lan Jiang, Jean-François Silvain, and Yongfeng Lu
Opt. Express 25(22) 27000-27007 (2017)