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Novel wavelength-resolved fluorescence detection for a high-throughput capillary electrophoresis system under a diascopic configuration.

Journal of chromatography. A (2008-04-15)
Shi-Wei Lin, Guan-Liang Chang, Che-Hsin Lin
ABSTRACT

This paper presents a novel method regarding a wavelength-resolved fluorescence detection scheme for high-throughput analysis of bio-samples in a micro-CE chip. Instead of using the conventional laser-induced fluorescence (LIF) microscope equipped with delicate spatial filters and complex control systems, this study adopts a hollow cone illumination generated using a dark-field condenser for exciting fluorescence in the microchannel and an ultraviolet-visible-near-infrared (UV-Vis-NIR) spectrometer for detecting the emission signals. Experimental results show that the proposed system is feasible for simultaneously detecting a mixed sample composed of Atto 610, Rhodamine B and fluorescein isothiocyanate (FITC) fluorescent dyes in a single test run. Furthermore, a mixed bio-sample composed of two mixed 16-mer single-stranded DNAs labeled with Cy3 and FITC fluorescent dyes is also successfully detected with the proposed system. The measured limit of detection (LOD) for detecting FITC of the proposed system can be as low as 5.4x10(-6)M (S/N=3). This proposed detection method has shown its potential on RNA identification and DNA sequencing applications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Atto 610-Biotin, BioReagent, suitable for fluorescence, ≥85% (HPCE)
Sigma-Aldrich
Atto 610 maleimide, BioReagent, suitable for fluorescence, ≥90% (coupling to thiols)
Sigma-Aldrich
Atto 610-Streptavidin, BioReagent, suitable for fluorescence

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