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ACS Chem Neurosci. 2014 May 21;5(5):329-34. doi: 10.1021/cn5000138. Epub 2014 Mar 31.

Label-free dopamine imaging in live rat brain slices.

Author information

1
Tata Institute of Fundamental Research , Homi Bhabha Road, Colaba, Mumbai 400005, India.

Abstract

Dopaminergic neurotransmission has been investigated extensively, yet direct optical probing of dopamine has not been possible in live cells. Here we image intracellular dopamine with sub-micrometer three-dimensional resolution by harnessing its intrinsic mid-ultraviolet (UV) autofluorescence. Two-photon excitation with visible light (540 nm) in conjunction with a non-epifluorescent detection scheme is used to circumvent the UV toxicity and the UV transmission problems. The method is established by imaging dopamine in a dopaminergic cell line and in control cells (glia), and is validated by mass spectrometry. We further show that individual dopamine vesicles/vesicular clusters can be imaged in cultured rat brain slices, thereby providing a direct visualization of the intracellular events preceding dopamine release induced by depolarization or amphetamine exposure. Our technique opens up a previously inaccessible mid-ultraviolet spectral regime (excitation ~270 nm, emission < 320 nm) for label-free imaging of native molecules in live tissue.

PMID:
24661118
PMCID:
PMC4030794
DOI:
10.1021/cn5000138
[Indexed for MEDLINE]
Free PMC Article

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