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Updated: Sep 10, 2025
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Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina
Published on: February 13, 2021
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Color-neutral and reversible tissue transparency enables longitudinal deep-tissue imaging in live mice.
Carl H C Keck1,2, Elizabeth L Schmidt1,2,3, Richard H Roth4
1Department of Materials Science and Engineering, Stanford University, Stanford, CA 94305.
Summary
Researchers developed a novel method for achieving optical transparency in live mice across the visible spectrum. This technique enables deep in vivo imaging and long-term tracking of cellular activity without causing harm.
Area of Science:
- Biomedical Optics
- In Vivo Imaging
- Tissue Engineering
Background:
- Light scattering in biological tissues impedes deep in vivo imaging.
- Previous methods achieved red spectrum transparency using absorbing dyes.
- Need for broader spectral transparency for diverse imaging applications.
Purpose of the Study:
- To extend optical transparency across the entire visible spectrum in live mice.
- To enable noninvasive deep-tissue imaging of cellular structures and dynamics.
- To facilitate longitudinal studies of biological processes in vivo.
Main Methods:
- Utilized molecules with strong ultraviolet absorption and sharp visible spectrum edges.
- Achieved color-neutral, reversible tissue transparency.
- Applied method for imaging yellow fluorescent protein and GCaMP in mouse brains through intact scalp and skull.
Main Results:
- Demonstrated optical transparency across the visible spectrum.
- Enabled structural and functional imaging of the mouse brain, including longitudinal studies over multiple days.
- Histological and toxicological studies showed minimal acute or chronic damage.
Conclusions:
- This color-neutral, reversible tissue transparency technique enhances deep-tissue optical imaging capabilities.
- Opens opportunities for noninvasive, long-term visualization of cellular activity with high spatiotemporal resolution.
- Facilitates chronic tracking of biological processes in live animal models.
