Random-access scanning microscopy for 3D imaging in awake behaving animals

Nat Methods. 2016 Dec;13(12):1001-1004. doi: 10.1038/nmeth.4033. Epub 2016 Oct 17.

Abstract

Understanding how neural circuits process information requires rapid measurements of activity from identified neurons distributed in 3D space. Here we describe an acousto-optic lens two-photon microscope that performs high-speed focusing and line scanning within a volume spanning hundreds of micrometers. We demonstrate its random-access functionality by selectively imaging cerebellar interneurons sparsely distributed in 3D space and by simultaneously recording from the soma, proximal and distal dendrites of neocortical pyramidal cells in awake behaving mice.

MeSH terms

  • Action Potentials / physiology
  • Animals
  • Behavior, Animal / physiology*
  • Cerebellar Cortex / cytology
  • Cerebellar Cortex / physiology
  • Dendrites / physiology
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • Imaging, Three-Dimensional / methods*
  • Interneurons / physiology
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Motor Activity / physiology*
  • Neurons / physiology*
  • Patch-Clamp Techniques
  • Pyramidal Cells / physiology
  • Visual Cortex / cytology
  • Visual Cortex / physiology
  • Voltage-Sensitive Dye Imaging / methods*

Substances

  • Green Fluorescent Proteins