Imaging large-scale neural activity with cellular resolution in awake, mobile mice

Neuron. 2007 Oct 4;56(1):43-57. doi: 10.1016/j.neuron.2007.08.003.


We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to approximately 2-5 microm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of <5%.

Publication types

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

MeSH terms

  • Algorithms
  • Animals
  • Astrocytes / physiology
  • Behavior, Animal
  • Brain / cytology*
  • Diagnostic Imaging / instrumentation
  • Diagnostic Imaging / methods*
  • Equipment Design
  • Exercise Test / methods
  • Female
  • Luminescent Proteins / metabolism
  • Male
  • Markov Chains
  • Mice
  • Mice, Transgenic
  • Models, Neurological
  • Motor Activity / physiology*
  • Neurons / physiology*
  • Organic Chemicals / metabolism
  • Wakefulness / physiology*


  • Luminescent Proteins
  • Organic Chemicals
  • calcium green