Raman microscopy unmixed: Technical advances, bio-orthogonal tags, and applications in life sciences

Raman microscopy has emerged as a transformative tool in chemical biology, enabling non-invasive, label-free visualization of biological processes with molecular specificity. This review aims to provide an overview of current Raman microscopy techniques—spanning spontaneous and coherent Raman modalities—and their applications in bioimaging. We outline the core technological advancements driving these techniques, including improvements in spatial, temporal, and spectral resolution, as well as innovations in multiplex imaging and volumetric capabilities. Aspiring to bridge gaps between spectroscopists and biologists, this review highlights the critical applications of Raman microscopy, such as cell metabolism studies, disease progression diagnostics, and tissue mapping, while also exploring emerging technologies like quantum light sources and time-resolved Raman microscopy. Ultimately, this review seeks to foster an appreciation for the evolving capabilities of Raman systems and their potential to enable unprecedented insight into complex biological processes, promoting cross-disciplinary collaboration and innovation in chemical biology.