Live-cell microscopy demands high specificity, sensitivity, and minimal perturbation to the biomolecules of interest. Meeting all these criteria has been challenging in cellular imaging. Toward this goal, a bioorthogonal Raman imaging method has recently emerged by exploiting small Raman reporters that possess Raman signals that do not overlap with the naturally existing biomolecules in the cells. The Raman reporters are metabolically incorporated into the target biomolecules for direct visualization. Herein, we review recent advances in the methodological development and the proof-of-concept applications of the live-cell bioorthogonal Raman imaging technique.
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