Large-scale deep tissue voltage imaging with targeted illumination confocal microscopy
Date
2023-07-21
Authors
Xiao, Sheng
Cunningham, William J.
Kondabolu, Krishnakanth
Lowet, Eric
Moya, Maria V.
Mount, Rebecca
Ravasio, Cara
Economo, Michael N.
Han, Xue
Mertz, Jerome
Version
First author draft
OA Version
Citation
S. Xiao, W.J. Cunningham, K. Kondabolu, E. Lowet, M.V. Moya, R. Mount, C. Ravasio, M.N. Economo, X. Han, J. Mertz. 2023. "Large-scale deep tissue voltage imaging with targeted illumination confocal microscopy." bioRxiv. https://doi.org/10.1101/2023.07.21.548930
Abstract
Voltage imaging with cellular specificity has been made possible by the tremendous advances in genetically encoded voltage indicators (GEVIs). However, the kilohertz rates required for voltage imaging lead to weak signals. Moreover, out-of-focus fluorescence and tissue scattering produce background that both undermines signal-to-noise ratio (SNR) and induces crosstalk between cells, making reliable in vivo imaging in densely labeled tissue highly challenging. We describe a microscope that combines the distinct advantages of targeted illumination and confocal gating, while also maximizing signal detection efficiency. The resulting benefits in SNR and crosstalk reduction are quantified experimentally and theoretically. Our microscope provides a versatile solution for enabling high-fidelity in vivo voltage imaging at large scales and penetration depths, which we demonstrate across a wide range of imaging conditions and different GEVI classes.
Description
License
The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a Attribution-NonCommercial-NoDerivs 4.0 International.