Wide field-of-view, multi-region, two-photon imaging of neuronal activity in the mammalian brain

Nat Biotechnol. 2016 Aug;34(8):857-62. doi: 10.1038/nbt.3594. Epub 2016 Jun 27.


Two-photon calcium imaging provides an optical readout of neuronal activity in populations of neurons with subcellular resolution. However, conventional two-photon imaging systems are limited in their field of view to ∼1 mm(2), precluding the visualization of multiple cortical areas simultaneously. Here, we demonstrate a two-photon microscope with an expanded field of view (>9.5 mm(2)) for rapidly reconfigurable simultaneous scanning of widely separated populations of neurons. We custom designed and assembled an optimized scan engine, objective, and two independently positionable, temporally multiplexed excitation pathways. We used this new microscope to measure activity correlations between two cortical visual areas in mice during visual processing.

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
  • Brain Mapping / instrumentation*
  • Brain Mapping / methods
  • Image Enhancement / instrumentation
  • Image Enhancement / methods
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Fluorescence, Multiphoton / instrumentation*
  • Microscopy, Fluorescence, Multiphoton / methods
  • Nerve Net / physiology
  • Neurons / cytology
  • Neurons / physiology*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Visual Cortex / physiology*
  • Visual Perception / physiology*
  • Voltage-Sensitive Dye Imaging / instrumentation*
  • Voltage-Sensitive Dye Imaging / methods