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Updated: May 9, 2026

Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain

Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain

Published on: January 13, 2022

Simultaneous three-photon and optical coherence microscopy deep within an intact mouse brain.

Xusan Yang^1,2, Siyang Liu³, Fei Xia^4,5,6

¹School of Applied and Engineering Physics, Cornell University, Ithaca, NY, USA. xusanyang@iphy.ac.cn.

|May 7, 2026

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This study combines three-photon microscopy (3PM) and spectral-domain optical coherence microscopy (SD-OCM) for deep tissue imaging. The multimodal approach allows simultaneous visualization of structures in the mouse brain in vivo.

Area of Science:

Biomedical Engineering
Neuroscience
Optical Imaging

Background:

Multimodal microscopy offers complementary tissue information in a single session.
Combining imaging techniques enhances visualization capabilities for complex biological samples.

Purpose of the Study:

To demonstrate a multimodal imaging approach combining three-photon microscopy (3PM) and spectral-domain optical coherence microscopy (SD-OCM).
To showcase the simultaneous use of an optical parametric amplifier (OPA) laser source for 3PM, third harmonic generation (THG), and SD-OCM.
To evaluate the system's performance for deep tissue imaging in vivo.

Main Methods:

Utilized a multimodal microscopy system integrating 3PM and SD-OCM.
Employed an optical parametric amplifier (OPA) laser source at a 1620 nm center wavelength.

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Last Updated: May 9, 2026

Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain

Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain

Published on: January 13, 2022

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Published on: October 8, 2013

Multiphoton Microscopy of Cleared Mouse Brain Expressing YFP

Multiphoton Microscopy of Cleared Mouse Brain Expressing YFP

Published on: September 23, 2012

Performed in vivo imaging of deep mouse brains.

Main Results:

Achieved simultaneous 3PM, THG, and SD-OCM imaging using a single OPA laser source.
Visualized fine structures like myelinated axons, neurons, and fiber tracts at depths exceeding 1 mm in adult mouse brains.
Demonstrated noninvasive imaging with high spatial resolution using combined linear and nonlinear contrast mechanisms.

Conclusions:

Simultaneous OCM and 3PM at long wavelengths provide a powerful tool for deep tissue imaging in vivo.
The demonstrated multimodal system offers a convenient and effective approach for neuroscience research.
This technique advances noninvasive visualization of neural structures in the intact brain.