Development of Raman probes for live cell tracking

Fast-acquisition Raman methodologies based on either coherent anti-Stokes Raman scattering (CARS) or stimulated Raman scattering (SRS), generate image contrast using the Raman active vibrational frequency of a given chemical, providing information on the biochemical composition of tissues allowing species-specific label-free visualisation. In addition, generation of bioorthogonal Raman probes which target the silent region of the Raman spectra of cells provide dramatic enhancements in contrast and signal-to-noise ratios. This increased Raman activity coupled with the ability to design spectroscopically distinct Raman species opens up the possibility of tracking individual cell populations in complex 3D environments with high temporal and spatial resolution and specificity. Using in silico DFT studies, the student will design a suite of novel highly Raman-active water-soluble tags to give a tunable pallet of vibrational frequencies with well-defined intracellular characteristics, which can be targeted to specific organelles. Murine T cells will be labelled with the Raman tags and their ability to infiltrate 3D tumour organoids monitored over time and their spatial distribution mapped. SRS and Hyperspectral CARS will allow combinatorial cell/organoid-labelling to be explored, and changes in organoid morphology, structure and viability to be monitored in real time without the use of fluorescent probes. Cross-system assessment of different imaging systems: including the custom-built platforms in Edinburgh and Cardiff (hyperspectral CARS, SRS, TPF), and the commercial Leica system at GSK, will provide information on the translation of the technology to pre-clinical applications.