| 光谱学与光谱分析 |
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| Study on Multi-Photon Excited Fluorescence Combined with Capillary Electrophoresis |
| SUN Yun-xu, ZHU Feng, MA Wan-yun* |
| Key Laboratory for Atomic and Molecular Nanosciences of Education Ministry, Department of Physics, Tsinghua University, Beijing 100084, China |
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Abstract This paper describes a new method, multiphoton excitation fluorescence detection combined with capillary electrophoresis separation. The excitation source was a self mode-locked femtosecond titanium-sapphire laser (Spectra-physics Inc.), producing a stream of pulses with a pulse duration of about 100 fs at 82 MHz repetition rate. Its average power was about 200 mW at 750 nm. The laser beam was focused into a thin wall flow cell of capillary electrophoresis. A high numerical aperture objective (100×NA 1.25) was chosen to focus the laser beam and to collect the fluorescence emitted by detecting molecules. Then the fluorescence was detected by a fast response PMT. All data were acquired and processed by a microcomputer. For three biological molecules, 5HT, FAD and NADH, it was demonstrated that they can be separated and detected efficiently by this method with three-photon and two-photon excitation respectively using only one 750 nm laser beam. The detection limits were 1.0×10-６ mol·L-1 for 5HT, 7.4×10-７ mol·L-1 for FAD and 9.8×10-７ mol·L-1 for NADH using the criterion of three standard deviations above background. The results are much better than those with UV absorption detection by one or two magnitudes.
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Received: 2003-06-01
Accepted: 2003-09-16
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Corresponding Authors:
MA Wan-yun
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| Cite this article: |
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SUN Yun-xu,ZHU Feng,MA Wan-yun. Study on Multi-Photon Excited Fluorescence Combined with Capillary Electrophoresis [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2005, 25(04): 502-505.
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| URL: |
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https://www.gpxygpfx.com/EN/Y2005/V25/I04/502 |
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