Calcium imaging of neural circuits with extended depth-of-field light-sheet microscopy

Opt Lett. 2016 Mar 1;41(5):855-8. doi: 10.1364/OL.41.000855.

Abstract

Increasing the volumetric imaging speed of light-sheet microscopy will improve its ability to detect fast changes in neural activity. Here, a system is introduced for brain-wide imaging of neural activity in the larval zebrafish by coupling structured illumination with cubic phase extended depth-of-field (EDoF) pupil encoding. This microscope enables faster light-sheet imaging and facilitates arbitrary plane scanning-removing constraints on acquisition speed, alignment tolerances, and physical motion near the sample. The usefulness of this method is demonstrated by performing multi-plane calcium imaging in the fish brain with a 416×832×160 μm field of view at 33 Hz. The optomotor response behavior of the zebrafish is monitored at high speeds, and time-locked correlations of neuronal activity are resolved across its brain.

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 / cytology
  • Caenorhabditis elegans
  • Calcium / metabolism*
  • Light*
  • Microscopy / methods*
  • Molecular Imaging
  • Neurons / metabolism*

Substances

  • Calcium