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Confocal Imaging of Fast Flash Photolysis of Caged Compounds in Cultured Neurons

  • Eduard Korkotian
  • Menahem SegalEmail author
Protocol
Part of the Neuromethods book series (NM, volume 152)

Abstract

The enhanced ability to visualize small neuronal compartments in live tissue, such as individual dendritic spine, is accompanied in recent years by a need for a precise, high temporal and spatial resolution ability to activate or suppress electrophysiological as well as biochemical properties within such compartments. Parallel rapid progress in molecular, cellular, and physics methodologies enabled the recruitment of novel technologies to the analysis of a wide spectrum of issues, from long lasting imaging of subcellular compartments in vitro as well as in vivo, to simultaneous recording of activities of networks of hundreds of neurons in behaving animals. In the present review, we will focus on a fast UV flash (4 ns) photolysis of caged molecules in a small sphere (<1 μm) near or within dendritic spines of cultured neurons. This method is faster and cheaper than the commonly used 2-photon uncaging, which requires a large investment in complex equipment. Our method can best be used with two-dimensional networks of neurons, grown in culture.

Key words

Uncaging Confocal microscope Dendritic spines Calcium 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of NeurobiologyThe Weizmann InstituteRehovotIsrael

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