Advancing live cell phenotyping for cancer and developmental biology via label-free Raman spectroscopy and phase tomography

Quantitative, rapid, and non-perturbative phenotyping of live cells is a critical unmet need across the life sciences. Conventional methods such as immunohistochemistry and omics require extensive sample manipulation, yield primarily qualitative data, and hinder further downstream analysis and applications. In contrast, label-free optical imaging offers non-invasive and quantitative access to phenotypic traits while preserving cell viability and sterility. We combine tomographic phase microscopy (TPM) with spontaneous Raman (SR) spectroscopy to enable comprehensive morpho-molecular profiling of live cells and cell colonies. This high-content method discriminates subtle phenotypic differences using fewer biological replicates, reducing both time and cost, as we demonstrate in live cancer cells at varying disease stages. To highlight the biocompatibility and sensitivity of our tools even for fragile, three-dimensional biological models, we observed live embryonic colonies during early development. We revealed the subtle morpho-molecular manifestation of pluripotency exit, the first fate decision event in embryonic growth.