Researcher Phd Student
CurrentProject title: ‘Towards photoacoustic neuroimaging in mice: a chemigenetic approach for acoustogenic calcium sensors’Developing novel photoacoustic labels and calcium sensors optimised for this modality, with the overall goal of achieving whole mouse brain in vivo imaging.Our approach uses chemigenetic reporters, combining quenched rhodamine derivatives (presenting low quantum yields and high NIR absorption) with the self-labelling HaloTag protein to provide labelling specificity. These acoustogenic ligands can generate calcium-dependent photoacoustic signal when bound to HaloCaMP, a calcium sensitive derivative of HaloTag.Key stages: • Synthesis of quenched rhodamine dyes and in vitro characterisation of their photo- and biophysical properties with and without HaloTag protein. Systematic rational chemical modifications were made to optimise these properties.• Assessment of in vivo labelling efficiency in mice• Engineering of the calcium-sensitive HaloTag-based protein scaffold.• Proof of principle PA imaging: tissue phantoms, cultured neurons, brain slices, ex vivo mouse brains.Lab techniques: organic synthesis and characterisation, UV-Vis and fluorescence spectroscopy, photoacoustic spectroscopy, phantom preparation; molecular cloning, protein expression and purification.Achievement: development of the first chemigenetic probes and calcium sensors for photoacoustic imaging.
