Large-scale deep tissue voltage imaging with targeted-illumination confocal microscopy

Sheng Xiao*, William J. Cunningham, Krishnakanth Kondabolu, Eric Lowet, Maria V. Moya, Rebecca A. Mount, Cara Ravasio, Emma Bortz, Dana Shaw, Michael N. Economo, Xue Han, Jerome Mertz

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

Voltage imaging with cellular specificity has been made possible by advances in genetically encoded voltage indicators. 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 the signal-to-noise ratio 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 signal-to-noise ratio 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 genetically encoded voltage indicator classes.

Original languageEnglish
Pages (from-to)1094-1102
Number of pages9
JournalNature Methods
Volume21
Issue number6
DOIs
Publication statusPublished - 5 Jun 2024

Bibliographical note

Publisher Copyright:
© The Author(s), under exclusive licence to Springer Nature America, Inc. 2024.

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