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Analysis of serotonin release from single neuron soma using capillary electrophoresis and laser-induced fluorescence with a pulsed deep-UV NeCu laser

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Abstract

The use of capillary electrophoresis (CE) with laser-induced fluorescence excited by ultraviolet (UV) lasers in the range 200–300 nm has been restricted by the available wavelengths and expense of UV lasers. The integration of a NeCu deep UV laser operating at 248.6 nm with a single channel CE system with post-column sheath flow detection allows detection limits for serotonin and tryptophan of 3.9×10-8 M and 4.5×10-8 M respectively. Single cell analysis of serotonergic metacerebral cells from the sea slug Aplysia californica yields a value of 800±85 fmol of serotonin in each cell soma. For the first time, serotonin is directly detected in electrically stimulated release from single metacerebral cell soma, with approximately 4% of the serotonin contained in the soma released from a semi-intact preparation with a 2 min electrical stimulation.

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Acknowledgements

This work was supported by the National Science Foundation (CHE-98–77071) and NIH (MH60261). The authors appreciate the advice and assistance of William Hug and Ray Raid (Photon System Inc.) concerning the operation of the NeCu laser, Xin Zhang (UIUC) for advice on 5-HT measurements, and Sarah Sheeley (UIUC) for assistance with construction of the CE system. A. californica were provided by the NCRR National Resource for Aplysia at the University of Miami.

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  1. Department of Chemistry and the Beckman Institute, University of Illinois, Urbana, 61801, USA

    Hai Miao, Stanislav S. Rubakhin & Jonathan V. Sweedler

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  1. Hai Miao
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  2. Stanislav S. Rubakhin
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  3. Jonathan V. Sweedler
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Correspondence to Jonathan V. Sweedler.

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Miao, H., Rubakhin, S.S. & Sweedler, J.V. Analysis of serotonin release from single neuron soma using capillary electrophoresis and laser-induced fluorescence with a pulsed deep-UV NeCu laser. Anal Bioanal Chem 377, 1007–1013 (2003). https://doi.org/10.1007/s00216-003-2191-8

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  • DOI: https://doi.org/10.1007/s00216-003-2191-8

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