Chronic cellular imaging of entire cortical columns in awake mice using microprisms

Neuron. 2013 Nov 20;80(4):900-13. doi: 10.1016/j.neuron.2013.07.052. Epub 2013 Oct 17.


Two-photon imaging of cortical neurons in vivo has provided unique insights into the structure, function, and plasticity of cortical networks, but this method does not currently allow simultaneous imaging of neurons in the superficial and deepest cortical layers. Here, we describe a simple modification that enables simultaneous, long-term imaging of all cortical layers. Using a chronically implanted glass microprism in barrel cortex, we could image the same fluorescently labeled deep-layer pyramidal neurons across their entire somatodendritic axis for several months. We could also image visually evoked and endogenous calcium activity in hundreds of cell bodies or long-range axon terminals, across all six layers in visual cortex of awake mice. Electrophysiology and calcium imaging of evoked and endogenous activity near the prism face were consistent across days and comparable with previous observations. These experiments extend the reach of in vivo two-photon imaging to chronic, simultaneous monitoring of entire cortical columns.

Publication types

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

MeSH terms

  • Animals
  • Axons / physiology
  • Behavior, Animal / physiology
  • Calcium / physiology
  • Cerebral Cortex / cytology
  • Cerebral Cortex / physiology*
  • Data Interpretation, Statistical
  • Electrophysiological Phenomena
  • Female
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Neural Pathways / physiology
  • Neuroimaging / instrumentation*
  • Neuroimaging / methods
  • Neurons / physiology*
  • Photic Stimulation
  • Physical Stimulation
  • Presynaptic Terminals / physiology
  • Subcellular Fractions / physiology
  • Thalamus / physiology
  • Vibrissae / physiology
  • Wakefulness


  • Calcium