# Three-photon imaging of hippocampal neurogenesis through the intact mouse brain

**Authors:** Yusaku Hontani, Xin Su, Yicheng Wu, Diana Machado, Megumi Mizoguchi, John Darby Cole, Hansjörg Kasper, Philipp Bethge, Fritjof Helmchen, Sebastian Jessberger

PMC · DOI: 10.21203/rs.3.rs-8267806/v1 · 2025-12-19

## TL;DR

This paper introduces a non-invasive three-photon imaging technique to study brain cell development in live mice without damaging the brain.

## Contribution

The novel contribution is optimizing three-photon microscopy for longitudinal imaging of the hippocampus in intact mouse brains.

## Key findings

- Three-photon imaging reaches depths up to 1800 μm in the intact mouse brain.
- The method enables longitudinal observation of neural stem cells in the hippocampal dentate gyrus.
- The approach provides new insights into structural plasticity in deep brain regions without invasive procedures.

## Abstract

Multiphoton imaging allows for the visualization of structural and functional plasticity within the central nervous system. However, gaining optical access to deep brain structures, such as the hippocampal dentate gyrus (DG), requires invasive approaches, causing brain damage. Here we optimize three-photon (3P) microscopy to perform longitudinal imaging of the DG in the intact brain at unprecedented depth of up to 1800 μm. We apply this approach to follow the dynamics of neural stem cells (NSCs) in the adult and developing DG, allowing for novel insights into structural plasticity deep within the intact mouse brain.

## Linked entities

- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Diseases:** brain damage (MESH:D001925)
- **Species:** Mus musculus (house mouse, species) [taxon 10090]

## Figures

7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12776475/full.md

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Source: https://tomesphere.com/paper/PMC12776475