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A liquid core waveguide fluorescence detector for multicapillary electrophoresis applied to DNA sequencing in a
A Hanning1, J Westberg, J Roeraade
1Royal Institute of Technology, Department of Analytical Chemistry, Stockholm, Sweden.
Electrophoresis
|September 23, 2000
Summary
A novel laser-induced fluorescence detector uses total internal reflection within capillaries as waveguides for sensitive multicapillary electrophoresis. This technology enables efficient illumination and imaging, paving the way for high-throughput DNA sequencing.
Area of Science:
- Analytical Chemistry
- Biophysical Chemistry
- Spectroscopy
Background:
- Multicapillary electrophoresis (MCE) requires sensitive detection systems for high-throughput analysis.
- Existing fluorescence detectors face challenges in sensitivity, optical crosstalk, and scalability.
Purpose of the Study:
- To develop and characterize a new laser-induced fluorescence (LIF) detector for MCE.
- To demonstrate the detector's capability for DNA sequencing applications.
Main Methods:
- Utilizing total internal reflection (TIR) for waveguiding fluorescence within capillaries acting as liquid core waveguides (LCWs).
- Arranging capillaries in planar and 2D arrays for efficient laser illumination and end-on imaging.
- Employing a prism for wavelength dispersion and analyzing four-color DNA sequencing data.
Main Results:
- Achieved intercapillary optical crosstalk below 0.5% with high stray light rejection.
- Demonstrated successful application in four-color DNA sequencing using a 91-capillary array.
- The compact detector design shows potential for very large-scale multiplexing.
Conclusions:
- The developed LIF detector offers high sensitivity and low crosstalk for MCE.
- The waveguiding principle and compact imaging enable efficient and scalable fluorescence detection.
- This technology is well-suited for advanced applications like high-throughput DNA sequencing.