Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points
Department of Neurology, Baltimore VA Medical Center, University of Maryland School of Medicine
Digital micromirror devices (DMD) can generate complex patterns in time and space with which to control neuronal excitability. Issues relevant to the design, construction, and operation of DMD systems are discussed. Such a system enabled the demonstration of non-linear integration across distal dendritic branch points.
Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory Neurons
1SISSA, International School for Advanced Studies, 2Istituto di Biofisica, Consiglio Nazionale delle Ricerche, 3SISSA Unit, Italian Institute of Technology
Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds. Here, we show how to obtain patch-clamp recordings combined with photolysis of caged cAMP or caged Ca for the study of olfactory transduction in dissociated mouse olfactory sensory neurons.
Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development
ISOF - Bio Free Radicals, Consiglio Nazionale delle Ricerche
Radical-based biomimetic chemistry has been applied to building-up libraries necessary for biomarker development.
Monitoring Changes in the Intracellular Calcium Concentration and Synaptic Efficacy in the Mollusc Aplysia
1Fishberg Department of Neuroscience and Friedman Brain Institute, Mt. Sinai School of Medicine, 2Phase Five Communications Inc.
We demonstrate how changes in the intracellular free calcium concentration and synaptic efficacy can be simultaneously monitored in a ganglion preparation of Aplysia. We image intracellular calcium using a fluorescent dye, Calcium Orange, and induce and monitor synaptic transmission with sharp (intracellular) electrodes.
Two-Photon-Based Photoactivation in Live Zebrafish Embryos
Molecular Cell Biology, Weizmann Institute of Science
Multiphoton microscopy allows control of low energy photons with deep optical penetration and reduced phototoxicity. We describe the use of this technology for live cell labeling in zebrafish embryos. This protocol can be readily adapted for photo-induction of various light-responsive molecules.
Mapping Inhibitory Neuronal Circuits by Laser Scanning Photostimulation
1Department of Anatomy and Neurobiology, School of Medicine, University of California, Irvine, 2Department of Biomedical Engineering, School of Engineering, University of California, Irvine
This paper introduces an approach of combining laser scanning photostimulation with whole cell recordings in transgenic mice expressing GFP in limited inhibitory neuron populations. The technique allows for extensive mapping and quantitative analysis of local synaptic circuits of specific inhibitory cortical neurons.
Isolation and Genetic Manipulation of Adult Cardiac Myocytes for Confocal Imaging
Institute for Molecular Cell Biology, Universty of Saarland
Adult cardiac myocytes are primary cells that can be isolated from animal hearts and cultured for several days. Within this culture period adenoviral gene transfer can be used to express genetically encoded biosensors (GEBs) or fluorescent fusion proteins. Both approaches allow cellular investigations by means of confocal microscopy.
Historical View and Physiology Demonstration at the NMJ of the Crayfish Opener Muscle
Department of Biology, University of Kentucky
The opener muscle of the crayfish leg is presented for its historical importance and experimental versatility in muscle phenotype, synaptic physiology and plasticity.
