Reviews of bio-orthogonal probes in bioscience by stimulated Raman scattering microscopy

Stimulated Raman scattering (SRS) microscopy, is a nonlinear optical imaging method for visualizing chemical content based on molecular vibrational bonds, with high sensitivity, resolution, speed, and specificity. In the current review, we provided a comprehensive and critical review of the most recent developments in the field of SRS in combination with bio-orthogonal Raman tags or labels in bioscience. Firstly, we introduced the fundamentals of SRS microscopy and the theory principle of bio-orthogonal Raman tags. In particular, present the applications of each kind of bio-orthogonal Raman tags, including heavy water (D₂O), stable isotope probes (SIP), and triple-bonds tags. And shared our vision for the remaining challenges, research needs, and potential future breakthroughs for SRS technology lastly. We envision that the advanced SRS imaging and analysis will be a major force in future biological discovery.