Large-scale voltage imaging in behaving mice using targeted illumination

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...

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Bibliographic Details
Main Authors: Xiao, Sheng, Lowet, Eric, Gritton, Howard J, Fabris, Pierre, Wang, Yangyang, Sherman, Jack, Mount, Rebecca A, Tseng, Hua-an, Man, Heng-Ye, Straub, Christoph, Piatkevich, Kiryl D, Boyden, Edward S, Mertz, Jerome, Han, Xue
Other Authors: Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Format: Article
Language:English
Published: Elsevier BV 2023
Online Access:https://hdl.handle.net/1721.1/148692
Description
Summary: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.