Two-photon photostimulation and imaging of neural circuits

Nat Methods. 2007 Nov;4(11):943-50. doi: 10.1038/nmeth1105. Epub 2007 Oct 28.

Abstract

We introduce an optical method to stimulate individual neurons in brain slices in any arbitrary spatiotemporal pattern, using two-photon uncaging of MNI-glutamate with beam multiplexing. This method has single-cell and three-dimensional precision. By sequentially stimulating up to a thousand potential presynaptic neurons, we generated detailed functional maps of inputs to a cell. We combined this approach with two-photon calcium imaging in an all-optical method to image and manipulate circuit activity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / chemistry
  • Action Potentials / drug effects
  • Animals
  • Brain Mapping / instrumentation
  • Brain Mapping / methods*
  • Calcium Signaling / drug effects
  • Cytophotometry / instrumentation
  • Cytophotometry / methods*
  • Electrophysiology
  • Excitatory Postsynaptic Potentials / drug effects
  • Glutamates / metabolism
  • Glutamine / pharmacology
  • Indoles / chemistry
  • Indoles / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Fluorescence, Multiphoton / instrumentation
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Neocortex / cytology
  • Neocortex / drug effects
  • Neocortex / physiology
  • Nerve Net / cytology
  • Nerve Net / metabolism
  • Nerve Net / physiology*
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / physiology
  • Reproducibility of Results
  • Tetrodotoxin / pharmacology

Substances

  • 4-methoxy-7-nitroindolinyl-glutamate
  • Acetates
  • Glutamates
  • Indoles
  • mag-indo-1 AM
  • Glutamine
  • Indo-1 pentaacetoxymethyl ester
  • Tetrodotoxin