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
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.
| 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 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/39808 |
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