Detecting differential metabolic changes in breast cancer under methionine regulation using DO-SRS and TPF microscopy

Breast cancer is a diverse disease rife with numerous subtypes with material impact on prognoses. Current methods may lack accuracy or be cost and time prohibitive, but optical techniques such as Isotope Probed Stimulated Raman Scattering microscopy (ip-SRS), and two photon excitation fluorescence (TPEF) microscopy can reveal spatial biomolecular information useful in distinguishing cell subtypes and phenotypic states rapidly and accurately. In the present study, we used heavy water and L-methionine to probe the enzymatic incorporation during scavenging and de novo biosynthesis of macromolecules in MCF10A, MCF7, and MDA-MB-231 breast cancer cells with spontaneous Raman spectro-microscopy and SRS microscopy, as well as their effects on cellular respiration and organelle health by imaging the NADH and flavin pools and labeled organelles with TPEF. This will enhance diagnostic efficacy and illustrate specific biochemical effects of manipulated nutrition and targeted therapies.