Microscope for Multicolour Single Molecule Fluorescence Detection, Imaging, and (Cross-)Correlation Spectroscopy

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A single molecule fluorescence imaging and spectroscopy system will be built from commercial components to provide important new capabilities that will advance leading research and training programs with biomedical, biotechnological, environmental, and materials science themes. The main components include a multi-laser engine and beam combiner, a high-sensitivity EMCCD camera and three-colour image splitter, avalanche photodiodes, photon counting electronics, and an inverted microscope frame and optics, filters, and optomechanics needed for integrating components and assembling the full system. The proposed system will be unique in its capacity for multicolour single molecule measurements in both an imaging format with immobilized fluorophores and a non-imaging format with freely diffusing fluorophores. The proposed design offers both state-of-the-art capability and flexibility to support multiple users and applications. Specifically, the system will offer single molecule fluorescence imaging with up to three colours in parallel, fluorescence burst coincidence measurements with up to four parallel colour channels, fluorescence correlation spectroscopy (FCS), and up to four-colour fluorescence cross-correlation spectroscopy (FCCS). It is believed that the proposed system would be one-of-a-kind in Canada. The system will be used to develop ultra-sensitive biological assays, including assays for extremely rare genetic mutations and imaging probes for complex cellular processes; to study diffusion and chemical reactions in atmospheric aerosol particles; for high-resolution structural analysis of protein folding dynamics; and for characterization of biomolecular binding processes and the formation of multi-protein complexes. The system will be part of the Laboratory for Advanced Spectroscopy and Imaging Research (LASIR) at UBC. In addition to enabling cutting-edge research in multiple areas, the proposed system will enhance the research and training of more than 50 current graduate student, undergraduate student, and postdoctoral trainees, and more than 200 total trainees (projected) over the next six years.