Color-neutral and reversible tissue transparency enables longitudinal deep-tissue imaging in live mice
Light scattering in biological tissue presents a significant challenge for deep in vivo imaging. Our previous work demonstrated the ability to achieve optical transparency in live mice using intensely absorbing dye molecules, which created transparency in the red spectrum while blocking shorter-wavelength photons. In this paper, we extend this capability to achieve optical transparency across the entire visible spectrum by employing molecules with strong absorption in the ultraviolet spectrum and sharp absorption edges that rapidly decline upon entering the visible spectrum. This color-neutral and reversible tissue transparency method enables optical transparency for imaging commonly used fluorophores in the green and yellow spectra. Notably, this approach facilitates tissue transparency for structural and functional imaging of the live mouse brain labeled with yellow fluorescent protein and GCaMP through the scalp and skull. We show that this method enables longitudinal imaging of the same brain regions in awake mice over multiple days during development. Histological analyses of the skin and systemic toxicology studies indicate minimal acute or chronic damage to the skin or body using this approach. This color-neutral and reversible tissue transparency technique opens opportunities for noninvasive deep-tissue optical imaging, enabling long-term visualization of cellular structures and dynamic activity with high spatiotemporal resolution and chronic tracking capabilities.
Authors
- Chonghe Wang (ORCID: https://orcid.org/0000-0002-1083-2666)
- Sa Cai (ORCID: https://orcid.org/0000-0002-1370-0028)
- Brendan M. Floyd (ORCID: https://orcid.org/0000-0001-7412-0492)
- Hermenegildo Garcı́a (ORCID: https://orcid.org/0000-0002-9664-493X)
- Mark L. Brongersma (ORCID: https://orcid.org/0000-0003-1777-8970)
- Tony Wyss‐Coray (ORCID: https://orcid.org/0000-0001-5893-0831)
- Andrew Park
- Peng Liao (ORCID: https://orcid.org/0000-0001-5740-7272)
- Kenneth Brinson (ORCID: https://orcid.org/0000-0002-1447-9169)
- Su Zhao (ORCID: https://orcid.org/0000-0001-5797-9190)
- Andy P. Tsai (ORCID: https://orcid.org/0000-0001-6400-544X)
- Guosong Hong (ORCID: https://orcid.org/0000-0002-8858-4471)
- Todd P. Coleman (ORCID: https://orcid.org/0000-0002-2144-2495)
- Gordon X. Wang
- Han Cui (ORCID: https://orcid.org/0000-0001-6473-9232)
- Richard H. Roth (ORCID: https://orcid.org/0000-0002-6855-999X)
- E. Schmidt (ORCID: https://orcid.org/0000-0002-7420-2997)
- Elyse Lopez
- Carl H. C. Keck (ORCID: https://orcid.org/0000-0003-1944-4165)
- Ani Baghdasaryan (ORCID: https://orcid.org/0000-0003-1324-5683)
- Jun Ding (ORCID: https://orcid.org/0000-0003-0690-1312)
- Kerriann M. Casey (ORCID: https://orcid.org/0000-0003-4228-928X)
- Qianru Yang (ORCID: https://orcid.org/0000-0003-4245-6631)
- Lauren Cooper (ORCID: https://orcid.org/0000-0003-3494-7609)
- Carolyn R. Bertozzi (ORCID: https://orcid.org/0000-0003-4482-2754)
- Miao Cui (ORCID: https://orcid.org/0009-0002-1413-7887)
- Xiaoyu Chen (ORCID: https://orcid.org/0000-0002-5764-2958)
- Lei Yuan (ORCID: https://orcid.org/0000-0001-7622-0809)
- Xiaoke Chen (ORCID: https://orcid.org/0000-0001-9032-9093)
- Wencke Reineking
- Daniel Esquivel
- Ling-Yi Zhang
Institutions
- Neurosciences Institute (US)
- Palo Alto Institute (US)
- Division of Materials Science and Engineering (AU)
- Bioengineering Center (RU)
- Stanford University (US)
Publication Details
- Journal
- Proceedings of the National Academy of Sciences
- Published
- 2025-08-26
- DOI
- https://doi.org/10.1073/pnas.2504264122
- Citations
- 2
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 3.27
Funders
- Damon Runyon Cancer Research Foundation
- National Institute on Aging
- National Institute of Neurological Disorders and Stroke
- Air Force Office of Scientific Research