A super-resolution microscope for visualizing cellular structure and dynamics

超分辨率共聚焦显微镜远低于光波长规定的分辨率极限，它违反了物理学定律，使细胞和发育生物学家能够俯视活细胞内正在工作的分子机器。这项在显微镜方面获得诺贝尔奖的进展正在迅速成为所有高影响分子细胞生物学研究的标准，包括西蒙弗雷泽大学(SFU)细胞生物学、发育和疾病中心(C2D2)的所有成员实验室进行的研究。除了对这种新的超分辨率技术的需求外，我们还迫切需要另一种多用途的共焦显微镜。目前，SFU只有两台共焦显微镜(覆盖整个校园)，通常可供183名教职员工和学生使用。除了超额认购外，这些共焦显微镜也是功能即将耗尽的老化系统。 我们建议获得一种基础型号的超分辨率共聚焦显微镜，它提供灵活性，既能满足SFU所有研究人员的常规显微镜需求，又能为我们机构提供其第一台超分辨率显微镜。配备徕卡Hyvolution的TCS SP8显微镜可以提供我们目前的共焦显微镜无法实现的巨大优势，即结合了高灵敏度、高速度、对任何样本都具有出色的穿透深度，以及最重要的是分辨率显著更高。除了购买设备所需的NSERC RTI外，对购买该系统的压倒性支持得到了许多承诺捐款，这些捐款来自SFU的三个部门(MBB、BISC、BPK)、两个学院(科学、卫生科学)和一些个人研究人员(主要申请者C.Beh博士和来自SFU的其他10名研究人员)。拟议系统的另一个好处是它是模块化的，允许今后对基础系统进行升级，以获得更高的分辨率能力。 建议的显微镜系统的灵活性与SFU分子细胞生物学实验室由NSERC资助的各种研究活动相匹配。作为带头申请者，BEH实验室将使用超分辨率显微镜直接显示内细胞膜之间的微小接触部位，以了解这些部位如何调节脂质和代谢物的来回运输。这10位共同申请者(T.Beischlag博士、S.Gorski博士、N.Harden博士、N.Hawkins博士、H.Hutter博士、M.Moore博士、G.Rintul博士、M.Silverman博士、G.Tibbit博士和D.vocadlo博士)将把新设备应用于各种研究领域，从神经细胞传输和调节器官发育的细胞过程，到癌症的细胞病因学和细胞内病原体的跟踪。作为向所有SFU用户开放的资源，受过培训的学生将从掌握这一技术先进的系统中受益，以便将来在生物技术领域就业。

Well below the resolution limit dictated by the wavelength of light, super-resolution confocal microscopes bend the laws of physics to allow cell and developmental biologist to peer down at molecular machines at work within living cells. This Nobel prize winning advance in microscopy is quickly becoming the standard for all high-impact molecular cell biology research, including that conducted by all the member labs of the Centre for Cell Biology, Development and Disease (C2D2) at Simon Fraser University (SFU). In addition to the need for this new super-resolution technology, we are also desperate for another multi-purpose confocal microscope. There currently exists only two confocal microscopes at SFU (for the entire campus) that are generally available to the >183 faculty and student users. Apart from being oversubscribed, these confocal microscopes are aging systems that are reaching the end of their functionality. We propose to acquire a base model super-resolution confocal microscope that offers flexibility to serve both the routine microscopy needs of all SFU researchers but also to provide our institution with its first super-resolution microscope. The TCS SP8 microscope with Hyvolution from Leica can provide tremendous benefits not fulfilled by our current confocal microscopes, namely, a combination of high sensitivity, speed, excellent depth of penetration for any sample, and most importantly, resolving power that is significantly higher. Overwhelming support for the acquisition of this system is underscored by the many pledged contributions, which are needed in addition to the NSERC RTI to purchase the equipment, coming from three SFU Departments (MBB, BISC, BPK), two Faculties (Science, Health Sciences), and a number of individual researchers (the lead applicant Dr. C. Beh and 10 other investigators from across SFU). The added benefit of the proposed system is that it is modular, permitting future upgrades to the base system for even higher resolution capabilities. The flexibility of the proposed microscope system matches the varied NSERC-funded research activities of the molecular cell biology labs at SFU. As the lead applicant, the Beh lab will use the super-resolution microscope to directly visualize the tiny contact sites between internal cellular membranes to understand how these sites mediate the transport of lipids and metabolites back-and-forth. The 10 co-applicants (Drs. T. Beischlag, S. Gorski, N. Harden, N. Hawkins, H. Hutter, M. Moore, G. Rintoul, M. Silverman, G. Tibbits, and D. Vocadlo) will apply the new equipment to various research interests ranging from nerve cell transport and cellular process regulating organ development, to cellular etiologies of cancer and the tracking of intracellular pathogens. As an open resource to all SFU users, trained students will benefit from the mastery of this technologically advanced system for future employment in biotech.