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Optical Clearing and Imaging of Immunolabeled Kidney Tissue
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Transparent tissue in solid state for solvent-free and antifade 3D imaging.

Fu-Ting Hsiao1, Hung-Jen Chien1, Ya-Hsien Chou1,2

  • 1Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan.

Nature Communications
|June 9, 2023
PubMed
Summary

Researchers developed a solid, high-refractive-index copolymer for 3D tissue imaging. This solvent-free method prevents evaporation and photobleaching, improving optical clearing and preserving fluorescent features for enhanced imaging and sharing.

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Area of Science:

  • Biomedical Engineering
  • Optical Imaging
  • Materials Science

Background:

  • Optical clearing is crucial for 3D tissue imaging, but current liquid reagents face challenges like solvent evaporation and photobleaching.
  • These issues compromise tissue optical properties and fluorescent signals, hindering detailed 3D visualization.

Purpose of the Study:

  • To develop a novel, stable clearing method for 3D tissue imaging.
  • To overcome limitations of liquid-based clearing agents, specifically evaporation and photobleaching.
  • To enable robust preservation, transfer, and sharing of cleared tissues for research and clinical applications.

Main Methods:

  • Designed a solid, high-refractive-index (high-n) acrylamide-based copolymer using the Gladstone-Dale equation as a guiding principle.
  • Embedded fluorescent dye-labeled mouse and human tissues within the solid copolymer matrix.
  • Utilized the solvent-free copolymer to minimize light scattering and dye fading during in-depth imaging.

Main Results:

  • The solid, high-n copolymer effectively cleared tissues without solvent evaporation or significant photobleaching.
  • Minimized light scattering in deep tissue imaging, preserving fluorescent signals.
  • Demonstrated a stable, liquid-free environment suitable for high/super-resolution 3D imaging.

Conclusions:

  • A novel solid, high-n copolymer offers a superior alternative to liquid-based clearing agents for 3D tissue imaging.
  • This solvent-free approach enhances tissue preservation, facilitates high-resolution imaging, and enables easier sharing of samples.
  • The developed method supports advanced investigation of tissue morphologies in both experimental and clinical settings.