With fast development and growing demand for cell-based immunotherapies and other immunomodulatory strategies e.g. microenvironment targeting, there is an urgent need to develop less invasive methods for cell analysis and quality control. Ideal tools serving these purposes should facilitate non-destructive, label-free analyses, retaining the analysed cell intact and unchanged. Raman spectroscopy (RS), measuring inelastic scattering corresponding to molecular vibration frequencies of different molecules, enables label-free subcellular imaging with high specificity and sensitivity. Through measuring metabolic state of the cells, RS may convey additional information e.g. regarding cell activation status, that can be utilized for diagnostic and research purposes.Recent success of chimeric antigen receptor (CAR) T cells and CAR-macrophages immunotherapies created a need for cell analysis methods allowing to identify activation status without affecting their therapeutic potential of cells. Since co-stimulated recognition of the antigen through the T cell receptor results in metabolic changes, clonal expansion, and cytokine production, T cell activation status is an ideal target for RS analysis. Similarly, macrophage phenotypes, namely pro-inflammatory M1 and immunosuppressive M2, are characterized by divergent metabolic status and potentially can be differentiated by means of RS.
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