Journal
JOURNAL OF NEUROSCIENCE METHODS
Volume 175, Issue 1, Pages 70-78Publisher
ELSEVIER
DOI: 10.1016/j.jneumeth.2008.08.003
Keywords
Caged compound; Uncaging; Optical fiber; Flash photolysis; MEA; Neuronal excitation
Categories
Funding
- Regione Lombardia [307727]
- MIUR [2006093522]
- Italian Institute of Technology
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Optical stimulation is a promising approach to investigate the local dynamic responses of cultured neurons. In particular, flash photolysis of caged compounds offers the advantage of allowing the rapid change of concentration of either extracellular or intracellular molecules, such as neurotransmitters or second messengers, for the stimulation or modulation of neuronal activity. We describe here the use of an ultraviolet (UV) laser diode coupled to an optical fibre for the local activation of caged compounds combined with a Micro-Electrode Array (MEA) device. Local uncaging was achieved by UV irradiation through the optical fibre previously positioned by using a red laser diode. The size of the stimulation was determined using caged fluorescein, whereas its efficacy was tested by studying the effect of uncaging the neurotransmitter glutamate. Uncaged glutamate evoked neuronal responses that were recorded using either fluorescence measurements or electrophysiological recordings with MEAs, thus showing the ability ofour system to induce local neuronal excitation. This method allows overcoming the limitations of the MEA system related to unfocused electrical stimulation and induction of electrical artefacts. In addition, the coupling of a UV laser diode to an optical fibre allows a precise local Stimulation and a quick change of the stimulation point. (C) 2008 Elsevier B.V. All rights reserved.
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