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

• Published in: Nature communications Vol. 14; no. 1; pp. 3395 - 12
• Main Authors: Hsiao, Fu-Ting, Chien, Hung-Jen, Chou, Ya-Hsien, Peng, Shih-Jung, Chung, Mei-Hsin, Huang, Tzu-Hui, Lo, Li-Wen, Shen, Chia-Ning, Chang, Hsiu-Pi, Lee, Chih-Yuan, Chen, Chien-Chia, Jeng, Yung-Ming, Tien, Yu-Wen, Tang, Shiue-Cheng
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 09.06.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 13/1; 13/51; 14/19; 14/34; 631/1647/245/2225; 631/1647/328/2238; 631/61/54/990; 692/698/1688/1315/1951; 692/698/2741/416; Acrylamide; Animals; Clearing; Copolymers; Dyes; Evaporation; Fluorescent Dyes; Fluorescent indicators; Human tissues; Humanities and Social Sciences; Humans; Image resolution; Imaging, Three-Dimensional - methods; Laboratory tests; Mice; multidisciplinary; Optical Imaging; Photobleaching; Photochemical reactions; Polymers; Reagents; Science; Science (multidisciplinary); Solid state; Solvents; Three dimensional imaging; Tissues
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-39082-4

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. Current liquid-based optical clearing protocols can suffer from solvent evaporation and photobleaching. Here, the authors develop a solid high-refractive-index polymer to embed mouse and human tissues for clearing and antifade high-resolution 3D imaging.