Induction and measurement of the early stage of a host-parasite interaction using a combined optical trapping and Raman microspectroscopy system.

Understanding and quantifying the temporal acquisition of host cell molecules by intracellular pathogens is fundamentally important in biology. In this study, a recently developed holographic optical trapping (HOT)-based Raman microspectroscopy (RMS) instrument is applied to detect, characterize and monitor in real time the molecular trafficking of a specific molecular species (isotope-labeled phenylalanine (L-Phe(D8)) at the single cell level. This approach enables simultaneous measurement of the chemical composition of human cerebrovascular endothelial cells and the protozoan parasite Toxoplasma gondii in isolation at the very start of the infection process. Using a model to decouple measurement contributions from host and pathogen sampling in the excitation volume, the data indicate that manipulating parasites with HOT coupled with RMS chemical readout was an effective method for measurement of L-Phe(D8) transfer from host cells to parasites in real-time, from the moment the parasite enters the host cell.