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

• Published in: Nature Communications
• Main Authors: 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, Shiue-Cheng Tang
• Format: Article
• Language: English
• Published: Nature Portfolio 2023-06-01
• Online Access: https://doi.org/10.1038/s41467-023-39082-4

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.