This research introduces RADIANT, a groundbreaking optical imaging method designed to map numerous proteins simultaneously within large, three-dimensional biological volumes. Conventional fluorescence microscopy is hindered by a "color barrier" that limits simultaneous imaging to a few targets, while existing high-multiplex methods are typically restricted to thin tissue sections. To overcome these obstacles, the authors utilize stimulated Raman scattering (SRS) microscopy combined with specially engineered MARS dyes, which feature narrow vibrational peaks that allow for the clear separation of many distinct signals. The study further develops rDISCO, a specialized tissue-clearing technique that enhances imaging depth by 10- to 100-fold while protecting the chemical stability of the Raman probes. By integrating these innovations, the researchers successfully demonstrated one-shot imaging of up to 12 targets in millimeter-thick brain tissues, providing a powerful tool for exploring complex 3D protein networks.

References:

• Shi, L., Wei, M., Miao, Y. et al. Highly-multiplexed volumetric mapping with Raman dye imaging and tissue clearing. Nat Biotechnol 40, 364–373 (2022). doi.org