Raman spectroscopy has been used to observe uptake, metabolism and response of single-cells to drugs. Photodynamic therapy is based on the use of light, a photosensitiser and oxygen to destroy tumour tissue. Here, we used single-cell Raman spectroscopy to study the uptake and intracellular degradation of a novel photosensitiser with a diphenylacetylene structure, DC473, in live single-cells from colorectal adenocarcinoma cell lines SW480, HT29 and SW620. DC473 was seen to predominantly accumulate in lipid droplets, showing higher accumulation in HT29 and SW620 cells than in SW480 cells, with a broader DC473 peak shifted to higher wavenumbers. DC473 activation and effects were tracked on live single-cells for 5 minutes. Upon exposure to UV light, the DC473 signal intensity dropped, with remaining DC473 shifting towards higher wavenumbers and widening, with a lifetime of approximately 50 seconds. Morphologically, SW480 and SW620 cells showed changes upon photodynamic therapy, whereas HT29 cells showed no changes. Morphological changes correlated with higher remaining DC473 signal after UV exposure. Our research suggests that DC473 forms aggregates within the cells that disaggregate following activation, showing the potential of Raman spectroscopy for the study of time-dependent single-cell pharmacodynamics.