Increasing Versatility in a Multi-User Laser Scanning Confocal Microscope

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The Olympus FV1000 laser scanning confocal microscope is used for 3-dimensional multi-color imaging that is essential to understand the spatial relationship of fluorescently tagged molecules and structures of interest in cells and tissues. This equipment is managed by the centralized UBC Bioimaging Facility and is heavily used: 22 HQP from 17 UBC research groups used this microscope in 2017. Funding is requested to upgrade the elements of the Olympus system that are failing and to increase the versality of live-cell long-term imaging. An upgrade to the current Olympus system would meet our urgent research needs for four reasons: to permit detection of yellow fluorescent proteins (YFP) (new laser), to sensitively detect fluorescent proteins (new detectors), to automatically collect images for compilation into large data sets (motorized stage and computer infrastructure), to increase the capability of live-cell long-term imaging (temperature control unit and silicon oil objective lenses). ******The Samuels lab has used this equipment to characterize the cells on cannabis flower that produce the valuable metabolites, but on-going studies are currently limited by the present detectors, computer infrastructure, and objective lenses. The Haughn lab uses YFP as a reporter to study the plant cell wall formation, but cannot presently use the system without a new YFP laser, detectors, and objective lenses for their live-cell imaging. The Auld lab studies the glial sheath formation in Drosophila melanogaster. A sensitive detector, new laser and temperature control unit are critical for their live-cell imaging of tagged proteins and calcium sensors. The Bohlmann lab requires the new laser, lenses, and detectors in their studies of tree defence by stone cells. The Wasteneys lab uses the Olympus system to perform Fluorescence Recovery after Photobleaching (FRAP) measurements to study microtubule dynamics and protein turn over controlling plant growth. The temperature control unit and the silicon oil lenses are essential for long-term imaging, e.g. during plant growth. The upgrade to the motorized stage will allow the tiling of whole roots to study epidermal cell file rotations during root growth. ******Upgrade of this equipment will benefit not only the co-applicants' labs, but many other research groups who require this type of live-cell imaging at UBC Vancouver. Due to the UBC Bioimaging Facility's dedicated technicians, HQP using this microscope receive excellent training and support. A delay in upgrading the Olympus system could negatively impact the thesis and research program of the 22 HQP relying on this microscope and would put this equipment at risk of becoming unusable.