Label-free dopamine imaging in live rat brain slices

Sarkar, Bidyut, Banerjee, Arkarup, Das, Anand Kant, Nag, Suman, Kaushalya, Sanjeev Kumar, Tripathy, Umakanta, Shameem, Mohammad, Shukla, Shubha, Maiti, Sudipta (May 2014) Label-free dopamine imaging in live rat brain slices. ACS Chemical Neuroscience, 5 (5). pp. 329-334. ISSN 1948-7193

DOI: 10.1021/cn5000138


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.

Item Type: Paper
Subjects: ?? Animals ??
?? Animals, Newborn ??
?? Brain ??
?? Cells, Cultured ??
?? Dopamine ??
?? In Vitro Techniques ??
?? Mass Spectrometry ??
?? Mice ??
?? Microscopy, Ultraviolet ??
?? Neuroglia ??
?? Neurons ??
?? Optical Imaging ??
?? Rats ??
?? Serotonin Agents ??
?? Time Factors ??
?? p-Chloroamphetamine ??
CSHL Authors:
Communities: CSHL labs > Banerjee lab
School of Biological Sciences > Publications
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 21 May 2014
Date Deposited: 03 Feb 2021 16:30
Last Modified: 01 Mar 2024 14:33
PMCID: PMC4030794
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