Video-rate volumetric neuronal imaging using 3D targeted illumination

Sci Rep. 2018 May 21;8(1):7921. doi: 10.1038/s41598-018-26240-8.


Fast volumetric microscopy is required to monitor large-scale neural ensembles with high spatio-temporal resolution. Widefield fluorescence microscopy can image large 2D fields of view at high resolution and speed while remaining simple and costeffective. A focal sweep add-on can further extend the capacity of widefield microscopy by enabling extended-depth-of-field (EDOF) imaging, but suffers from an inability to reject out-of-focus fluorescence background. Here, by using a digital micromirror device to target only in-focus sample features, we perform EDOF imaging with greatly enhanced contrast and signal-to-noise ratio, while reducing the light dosage delivered to the sample. Image quality is further improved by the application of a robust deconvolution algorithm. We demonstrate the advantages of our technique for in vivo calcium imaging in the mouse brain.

Publication types

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

MeSH terms

  • Algorithms*
  • Animals
  • Brain / diagnostic imaging
  • Brain / physiology*
  • Female
  • Fluorescence
  • Image Processing, Computer-Assisted / methods*
  • Lighting / instrumentation*
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence / methods*
  • Neuroimaging / methods*
  • Video Recording*