title: Spatially selective photoconductive stimulation of live neurons.
abstract: Synaptic activity is intimately linked to neuronal structure and function. Stimulation of live cultured primary neurons, coupled with fluorescent indicator imaging, is a powerful technique to assess the impact of synaptic activity on neuronal protein trafficking and function. Current technology for neuronal stimulation in culture include chemical techniques or microelectrode or optogenetic based techniques. While technically powerful, chemical stimulation has limited spatial resolution and microelectrode and optogenetic techniques require specialized equipment and expertise. We report an optimized and improved technique for laser based photoconductive stimulation of live neurons using an inverted confocal microscope that overcomes these limitations. The advantages of this approach include its non-invasive nature and adaptability to temporal and spatial manipulation. We demonstrate that the technique can be manipulated to achieve spatially selective stimulation of live neurons. Coupled with live imaging of fluorescent indicators, this simple and efficient technique should allow for significant advances in neuronal cell biology.
authors.affiliation: Developmental Neuroscience, Munroe-Meyer Institute, University of Nebraska Medical Center Omaha, NE, USA.
authors.forename: Jacob
authors.initials: J
authors.lastname: Campbell
authors.affiliation: Developmental Neuroscience, Munroe-Meyer Institute, University of Nebraska Medical Center Omaha, NE, USA.
authors.forename: Dipika
authors.initials: D
authors.lastname: Singh
authors.affiliation: Materials Science and Engineering Program, University of California San Diego, La Jolla, CA, USA.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Hollett
authors.affiliation: Department of Pharmacology, Creighton University Omaha, NE, USA.
authors.forename: Shashank M
authors.initials: SM
authors.lastname: Dravid
authors.affiliation: Materials Science and Engineering Program, University of California San Diego, La Jolla, CA, USA ; Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Sailor
authors.affiliation: Developmental Neuroscience, Munroe-Meyer Institute, University of Nebraska Medical Center Omaha, NE, USA.
authors.forename: Jyothi
authors.initials: J
authors.lastname: Arikkath
keywords: couple with live imaging; non-invasive neuronal stimulation; photoconductive stimulation; primary neurons; spatially selective stimulation