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A liquid core waveguide fluorescence detector for multicapillary electrophoresis applied to DNA sequencing in a 91‐capillary array
Author(s) -
Hanning Anders,
Westberg Joakim,
Roeraade Johan
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20000901)21:15<3290::aid-elps3290>3.0.co;2-j
Subject(s) - optics , capillary electrophoresis , detector , capillary action , laser , materials science , fluorescence , stray light , laser induced fluorescence , cylindrical lens , rhodamine , optoelectronics , chemistry , lens (geology) , physics , chromatography , composite material
A new laser‐induced fluorescence (LIF) detector for multicapillary electrophoresis is presented. The detection principle is based on waveguiding of the emitted fluorescence from the point of illumination to the capillary ends by total internal reflection (TIR) and imaging of the capillary ends. The capillaries themselves thus act as liquid core waveguides (LCWs). At the illumination point, the capillaries are arranged in a planar array, which allows clean and efficient illumination with a line‐focused laser beam. The capillary ends are rearranged into a small, densely packed two‐dimensional array, which is imaged end‐on with high light collection efficiency and excellent image quality. Wavelength dispersion is obtained with a single prism. Intercapillary optical crosstalk is less than 0.5%, and rejection of stray light is very efficient. The detector is applied to four‐color DNA sequencing by gel electrophoresis in a 91‐capillary array, with simple fluorescein and rhodamine dyes as fluorophores. Since the imaged two‐dimensional array is so compact, the detector has a high potential for very large‐scale multiplexing.