A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging

Aaron T. Mok; Tianyu Wang; Shitong Zhao; Kristine E. Kolkman; Danni Wu; Dimitre G. Ouzounov; Changwoo Seo; Chunyan Wu; Joseph R. Fetcho; Chris Xu

doi:10.1186/s43593-024-00076-4

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A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging

eLight 2024年4卷

RESEARCH ARTICLE | Updated：2025-04-15

- A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging
- A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging
- Affiliations：
- Author bio：
  
  atm624@cornell.edu
  bwangty@bu.edu
  kcx10@cornell.edu
- Funds：
  
  National Science Foundation;National Institute of Neurological Disorders and Stroke
- DOI：10.1186/s43593-024-00076-4
  中图分类号：
- 收稿日期：2024-06-23，
  
  修回日期：2024-08-28，
  
  录用日期：2024-09-20，
  
  网络出版日期：2024-11-01，
  
  纸质出版日期：2024-12
- Accepted：
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A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging[J]. eLight, 2024,4.

Aaron T. Mok, Tianyu Wang, Shitong Zhao, et al. A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging[J]. Elight, 2024, 4.
A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging[J]. eLight, 2024,4. DOI： 10.1186/s43593-024-00076-4.

Aaron T. Mok, Tianyu Wang, Shitong Zhao, et al. A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging[J]. Elight, 2024, 4. DOI： 10.1186/s43593-024-00076-4.

摘要

Abstract

In vivo imaging of large-scale neuronal activity plays a pivotal role in unraveling the function of the brain's circuitry. Multiphoton microscopy

a powerful tool for deep-tissue imaging

has received sustained interest in advancing its speed

field of view and imaging depth. However

to avoid thermal damage in scattering biological tissue

field of view decreases exponentially as imaging depth increases. We present a suite of innovations to optimize three-photon microscopy for large field-of-view imaging at depths unreachable by two-photon microscopy. These techniques enable us to image neuronal activities of transgenic animals expressing protein calcium sensors in a ~ 3.5-mm diameter field-of-view with single-cell resolution in the deepest cortical layer of mouse brains. We further demonstrate simultaneous large field-of-view two-photon and three-photon imaging

subcortical imaging in the mouse brain

and whole-brain imaging in adult zebrafish. The demonstrated techniques can be integrated into typical multiphoton microscopes to enlarge field of view for system-level neural circuit research.

关键词

Keywords

references

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