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Real-time cellular uptake of serotonin using fluorescence lifetime imaging with two-photon excitation

Botchway, SW, Parker, AW, Bisby, RH and Crisostomo, AG 2008, 'Real-time cellular uptake of serotonin using fluorescence lifetime imaging with two-photon excitation' , Microscopy Research and Technique, 71 (4) , pp. 267-273.

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Abstract

The real-time uptake of serotonin, a neurotransmitter, by rat leukaemia mast cell line RBL-2H3 and 5-hydroxytryptophan by Chinese hamster V79 cells has been studied by fluorescence lifetime imaging microscopy (FLIM), monitoring ultraviolet (340 nm) fluorescence induced by two-photon sub-picosecond 630 nm excitation. Comparison with two-photon excitation with 590 nm photons or by three-photon excitation at 740 nm (Williams et al, 1999) shows that the use of 630 nm excitation provides optimal signal intensity and lowered background from auto-fluorescence of other cellular components. In intact cells, we observe using FLIM three distinct fluorescence lifetimes of serotonin and 5-hydroxytryptophan according to location. The normal fluorescence lifetimes of both serotonin (3.8 ns) and 5-hydroxytryptophan (3.5 ns) in solution are reduced to ~2.5 ns immediately on uptake into the cell cytosol. The lifetime of internalized serotonin in RBL-2H3 cells is further reduced to ~2.0 ns when stored within secretory vesicles.

Item Type: Article
Uncontrolled Keywords: Serotonin; fluorescence; multiphoton; cellular; imaging; lifetime; microscopy; 5-hydroxytryptophan; FLIM
Themes: Subjects / Themes > Q Science > QH Natural history > QH301 Biology
Subjects outside of the University Themes
Schools: Colleges and Schools > College of Science & Technology
Colleges and Schools > College of Science & Technology > School of Environment and Life Sciences
Colleges and Schools > College of Science & Technology > School of Environment and Life Sciences > Biomedical Research Centre
Journal or Publication Title: Microscopy Research and Technique
Publisher: John Wiley & Sons
Refereed: Yes
ISSN: 1059910X
Funders: Science and Technology Facilities Council (STFC)
Depositing User: RH Bisby
Date Deposited: 24 Jun 2009 12:01
Last Modified: 17 Feb 2014 15:46
URI: http://usir.salford.ac.uk/id/eprint/2137

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