Large-scale voltage imaging in behaving mice using targeted illumination

Sheng Xiao, Eric Lowet, Howard J. Gritton, Pierre Fabris, Yangyang Wang, Jack Sherman, Rebecca A. Mount, Hua an Tseng, Heng Ye Man, Christoph Straub, Kiryl D. Piatkevich, Edward S. Boyden, Jerome Mertz, Xue Han

Research output: Contribution to journalArticlepeer-review


Recent improvements in genetically encoded voltage indicators enabled optical imaging of action potentials and subthreshold transmembrane voltage in vivo. To perform high-speed voltage imaging of many neurons simultaneously over a large anatomical area, widefield microscopy remains an essential tool. However, the lack of optical sectioning makes widefield microscopy prone to background cross-contamination. We implemented a digital-micromirror-device-based targeted illumination strategy to restrict illumination to the cells of interest and quantified the resulting improvement both theoretically and experimentally with SomArchon expressing neurons. We found that targeted illumination increased SomArchon signal contrast, decreased photobleaching, and reduced background cross-contamination. With the use of a high-speed, large-area sCMOS camera, we routinely imaged tens of spiking neurons simultaneously over minutes in behaving mice. Thus, the targeted illumination strategy described here offers a simple solution for widefield voltage imaging of many neurons over a large field of view in behaving animals.

Original languageEnglish (US)
Article number103263
Issue number11
StatePublished - Nov 19 2021


  • Neuroscience
  • Optical imaging
  • Techniques in neuroscience

ASJC Scopus subject areas

  • General


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