Two-photon Calcium Imaging From Head-Fixed Drosophila During Optomotor Walking Behavior

Nat Methods. 2010 Jul;7(7):535-40. doi: 10.1038/nmeth.1468. Epub 2010 Jun 6.


Drosophila melanogaster is a model organism rich in genetic tools to manipulate and identify neural circuits involved in specific behaviors. Here we present a technique for two-photon calcium imaging in the central brain of head-fixed Drosophila walking on an air-supported ball. The ball's motion is tracked at high resolution and can be treated as a proxy for the fly's own movements. We used the genetically encoded calcium sensor, GCaMP3.0, to record from important elements of the motion-processing pathway, the horizontal-system lobula plate tangential cells (LPTCs) in the fly optic lobe. We presented motion stimuli to the tethered fly and found that calcium transients in horizontal-system neurons correlated with robust optomotor behavior during walking. Our technique allows both behavior and physiology in identified neurons to be monitored in a genetic model organism with an extensive repertoire of walking behaviors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology
  • Brain / physiology
  • Calcium / metabolism*
  • Drosophila melanogaster / physiology*
  • Fluorescence
  • Green Fluorescent Proteins
  • Image Processing, Computer-Assisted / instrumentation*
  • Image Processing, Computer-Assisted / methods*
  • Motion
  • Motor Activity / physiology*
  • Neurons / physiology
  • Signal Transduction / physiology
  • Walking / physiology*


  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Calcium