AI Article Synopsis
- Mapping proteins in three-dimensional (3D) contexts is valuable in biomedicine, but existing fluorescence methods struggle with thick samples.
- The new technique, RADIANT, combines Raman dye imaging and tissue clearing to allow for simultaneous imaging of multiple targets in thick tissues, covering up to 11 different proteins in millimeter-thick brain slices.
- RADIANT enhances our understanding of protein interactions and networks, particularly in developmental processes like cerebellum formation, by extending imaging depth significantly compared to traditional methods.
Article Abstract
Mapping the localization of multiple proteins in their native three-dimensional (3D) context would be useful across many areas of biomedicine, but multiplexed fluorescence imaging has limited intrinsic multiplexing capability, and most methods for increasing multiplexity can only be applied to thin samples (<100 µm). Here, we harness the narrow spectrum of Raman spectroscopy and introduce Raman dye imaging and tissue clearing (RADIANT), an optical method that is capable of imaging multiple targets in thick samples in one shot. We expanded the range of suitable bioorthogonal Raman dyes and developed a tissue-clearing strategy for them (Raman 3D imaging of solvent-cleared organs (rDISCO)). We applied RADIANT to image up to 11 targets in millimeter-thick brain slices, extending the imaging depth 10- to 100-fold compared to prior multiplexed protein imaging methods. We showcased the utility of RADIANT in extracting systems information, including region-specific correlation networks and their topology in cerebellum development. RADIANT will facilitate the exploration of the intricate 3D protein interactions in complex systems.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8930416 | PMC |
| http://dx.doi.org/10.1038/s41587-021-01041-z | DOI Listing |
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