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[Study on multi-photon excited fluorescence combined with capillary electrophoresis].
- Source :
-
Guang pu xue yu guang pu fen xi = Guang pu [Guang Pu Xue Yu Guang Pu Fen Xi] 2005 Apr; Vol. 25 (4), pp. 502-5. - Publication Year :
- 2005
-
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 x 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 x 10(-6) mol x L(-1) for 5HT, 7.4 x 10(-7) mol x L(-1) for FAD and 9.8 x 10(-7) mol x 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.
Details
- Language :
- Chinese
- ISSN :
- 1000-0593
- Volume :
- 25
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Guang pu xue yu guang pu fen xi = Guang pu
- Publication Type :
- Academic Journal
- Accession number :
- 16097670