We are developing open multidimensional fluorescence microscopy instrumentation, including endomicroscopy, high content analysis (HCA), super-resolved microscopy, and optical projection tomography (OPT). We have particularly focused on fluorescence lifetime imaging (FLIM) and Forster resonant energy transfer (FRET) to study molecular interactions and more recently on super-resolved microscopy using single molecule localisation microscopy (SMLM) to probe ultrastructure and molecular clustering. To provide a complementary label-free readout, we are developing semi-quantitative (single-shot) phase contrast imaging for cell segmentation, tracking and morphology quantification.
For our current and future fluorescence microscopy, we are developing a modular open-source microscopy platform based on openFrame, a low-cost, modular, open microscopy hardware platform to be used with open-source software tools, including MicroManager and FIJI, for instrument control, data acquisition, analysis and management, in order to make them practical in lower resource settings. We are particularly focussing on implementing our techniques in an open-source HCA platform for more robust cell biology studies.
For FLIM/FRET HCA, we have developed an automated multiwell plate FLIM platform utilising open-source software for data acquisition and analysis, which we have applied to assay protein interactions, including applications to viral disease processes. For super-resolved HCA, we are developing automated multiwell plate easySTORM, providing low-cost, large FOV SMLM together with accelerated open-source SMLM analysis parallelised on a high-performance computing cluster. We have applied easySTORM in studies of defective phagocytosis, cancer and kidney disease.
For clinical applications we are developing histoSTORM – an implementation of easySTORM with frozen or FFPE tissue sections and clinically-approved antibody labelling. Other open microscopy developments include a low-cost modular OPT platform that can image mm-cm scale samples, including live zebrafish, and can provide single-shot volumetric imaging.