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 Vol. 4, (2024)

Research Article | Updated：2025-04-15

- 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：
- DOI：10.1186/s43593-024-00076-4
  CLC：
- Received：23 June 2024，
  
  Revised：28 August 2024，
  
  Accepted：2024-09-20，
  
  Published Online：01 November 2024，
  
  Published：2024-12
- Accepted：
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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：

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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