摘要：SummaryRecent improvements in genetically encoded voltage indicators enabled optical imaging of action potentials and subthreshold transmembrane voltagein 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.Graphical abstractDisplay OmittedHighlights•An easily implementable digital-micromirror-based targeted illumination microscope•Targeted illumination improves voltage imaging signal contrast in the brain•Targeted illumination decreases photobleaching and reduces background in the brain•Routine voltage imaging of tens of neurons simultaneously in behaving miceOptical imaging; Neuroscience; Techniques in neuroscience
