Transparent tissue in solid state for solvent-free and antifade 3D imaging

Fu-Ting Hsiao, Hung-Jen Chien, Ya-Hsien Chou, Shih-Jung Peng, Mei-Hsin Chung, Tzu-Hui Huang, Li-Wen Lo, Chia-Ning Shen, Hsiu-Pi Chang, Chih-Yuan Lee, Chien-Chia Chen, Yung-Ming Jeng, Yu-Wen Tien and Shiue-Cheng Tang ()
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Fu-Ting Hsiao: National Tsing Hua University
Hung-Jen Chien: National Tsing Hua University
Ya-Hsien Chou: National Tsing Hua University
Shih-Jung Peng: National Tsing Hua University
Mei-Hsin Chung: National Tsing Hua University
Tzu-Hui Huang: National Tsing Hua University
Li-Wen Lo: Academia Sinica
Chia-Ning Shen: Academia Sinica
Hsiu-Pi Chang: National Taiwan University Hospital
Chih-Yuan Lee: National Taiwan University Hospital
Chien-Chia Chen: National Taiwan University Hospital
Yung-Ming Jeng: National Taiwan University Hospital
Yu-Wen Tien: National Taiwan University Hospital
Shiue-Cheng Tang: National Tsing Hua University

Nature Communications, 2023, vol. 14, issue 1, 1-12

Abstract: Abstract Optical clearing with high-refractive-index (high-n) reagents is essential for 3D tissue imaging. However, the current liquid-based clearing condition and dye environment suffer from solvent evaporation and photobleaching, causing difficulties in maintaining the tissue optical and fluorescent features. Here, using the Gladstone-Dale equation [(n−1)/density=constant] as a design concept, we develop a solid (solvent-free) high-n acrylamide-based copolymer to embed mouse and human tissues for clearing and imaging. In the solid state, the fluorescent dye-labeled tissue matrices are filled and packed with the high-n copolymer, minimizing scattering in in-depth imaging and dye fading. This transparent, liquid-free condition provides a friendly tissue and cellular environment to facilitate high/super-resolution 3D imaging, preservation, transfer, and sharing among laboratories to investigate the morphologies of interest in experimental and clinical conditions.

Date: 2023
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DOI: 10.1038/s41467-023-39082-4

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