Increasing Versatility in a Multi-User Laser Scanning Confocal Microscope

Olympus FV 1000激光扫描共聚焦显微镜用于三维多色成像，这对于了解细胞和组织中荧光标记分子和感兴趣结构的空间关系至关重要。该设备由中央UBC生物成像设施管理，并被大量使用：2017年，来自17个UBC研究小组的22名HQP使用了该显微镜。要求提供资金，以更新Olympus系统中出现故障的部分，并提高活细胞长期成像的通用性。升级现有的奥林巴斯系统将满足我们紧迫的研究需求，原因有四：允许检测黄色荧光蛋白（YFP）（新激光），灵敏地检测荧光蛋白（新探测器），自动收集图像，用于汇编成大型数据集（机动舞台和计算机基础设施），提高活细胞长期成像的能力（温度控制单元和硅油物镜）。** 塞缪尔实验室已经使用这种设备来表征大麻花上产生有价值代谢物的细胞，但目前正在进行的研究受到现有检测器，计算机基础设施和物镜的限制。Haughn实验室使用YFP作为报告者来研究植物细胞壁的形成，但如果没有新的YFP激光器、探测器和物镜来进行活细胞成像，目前无法使用该系统。奥尔德实验室研究果蝇神经胶质鞘的形成。一个灵敏的探测器，新的激光器和温度控制单元对于标记蛋白质和钙传感器的活细胞成像至关重要。Bohlmann实验室需要新的激光器、透镜和探测器来研究石细胞对树木的防御。该实验室使用奥林巴斯系统进行光漂白后荧光恢复（FRAP）测量，以研究微管动力学和蛋白质周转控制植物生长。温度控制单元和硅油透镜对于长期成像至关重要，例如在植物生长期间。升级到机动阶段将允许平铺整个根，以研究根生长过程中的表皮细胞文件旋转。** 该设备的升级不仅有利于共同申请人的实验室，也有利于UBC温哥华需要这种活细胞成像的许多其他研究小组。由于UBC生物成像设施的专业技术人员，HQP使用这种显微镜接受良好的培训和支持。延迟升级奥林巴斯系统可能会对依赖该显微镜的22 HQP的论文和研究计划产生负面影响，并将使该设备处于无法使用的风险之中。

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.