Super-multiplexed vibrational imaging for 3D spatial biology
Understanding complex biological systems requires simultaneous characterization of a large number of interacting components in their native 3D environment. However, fluorescence methods are usually hindered by the fundamental color barrier. Here I will introduce electronic pre-resonance stimulated Raman scattering (epr-SRS) microscopy, an emerging method of ultrasensitive vibrational imaging with high potential of multiplexing. To generalize epr-SRS to large-scale volumetric imaging, we integrated it with tissue clearing technology and developed a technique named RADIANT. RADIANT achieved simultaneous visualization of >10 protein targets over millimeter thickness of brain tissues. Additionally, we bought multiplex protein imaging to nanoscopy with a new platform called MAGINIFIERS by combining epr-SRS with the recent advances of expansion microscopy. Overall, super-multiplexed vibrational imaging is a promising tool to provide a complete picture of tissue biology.
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