Acquisition of a super-resolution optical microscope

描述(由申请人提供)：本建议书要求提供资金，从应用精密公司购买新的超分辨率光学显微镜OMX Blaze V4，以加强范德比尔特大学NIH资助的主要用户的研究项目。成像技术的最新发展使得分辨生物样品中的对象成为可能，其尺寸小于传统光学显微镜的衍射极限(~200 nm)。从细胞生物学家的角度来看，这是一项关键的突破；蛋白质、蛋白质复合体、小泡和其他较小的细胞器都在200纳米以下。因此，超分辨率光学显微镜现在可以直接观察这些细微的亚细胞特征，甚至可以在完整的固定和活细胞的背景下精确定位单分子。应用精密公司的OMX blaze V4系统使用户能够利用两种超分辨率成像模式：结构照明显微镜(SIM)，用于快速同时对活细胞进行多色超分辨率成像(横向分辨率=80-100 nm)，以及定位显微镜(LM)，它为固定样本的成像提供尽可能高的横向分辨率(横向分辨率=20-50 nm)。主要用户(80%的使用率)是新成立的上皮生物学中心(EBC，由医学博士Bob Coffey和医学博士Jim Goldenring共同指导)的成员；这些研究人员将应用超分辨率成像技术，在他们由NIH资助的研究项目中开辟新的天地，这些项目涉及细胞和发育生物学、上皮生物学、癌症生物学和微生物学的广泛问题。项目包括研究细胞骨架动力学(Kiverina，Ohi，Tyska)，极化膜运输(Coffey，Crowe，Goldenring，Kiverina)，有丝分裂纺锤体的组装和功能(OHI)，细胞表面特征的组装，如微绒毛和纤毛(Goldenring，Tyska)，以及宿主与病原体的相互作用(Crowe，Tyska)。购买该系统还将使一组在亚蜂窝级别研究互补问题的小用户(使用率为20%)受益(Janetopoulos、Kenworth、Miller、Prince、Webb和Weaver)。总而言之，这些研究对于了解与出生缺陷、癌症、细菌/病毒感染和许多其他人类健康问题有关的疾病和疾病具有重要意义。拟议的OMX显微镜将安装在EBC中，并由Vanderbilt细胞成像共享资源(由Sam Wells，Ph.D.管理)管理。在范德比尔特研究办公室的大力支持下，该研究所将提供基础设施，向范德比尔特(约275个实验室)的研究人员介绍这项令人振奋的新技术，培训用户正确使用该仪器，并应用已建立的商业模式支持该系统的长期维护。总之，收购OMX将提供范德比尔特园区目前所不具备的前所未有的细胞成像能力。这一仪器将使研究人员能够以接近分子尺寸的空间分辨率为细胞成像，推动生物学问题的研究朝着目前范德比尔特用户可用的显微镜根本不可能的方向发展。

DESCRIPTION (provided by applicant): This proposal requests funds to purchase a new super-resolution optical microscope, the OMX Blaze V4 from Applied Precision, to enhance the NIH-funded research projects of Major Users at Vanderbilt University. Recent developments in imaging technology have made it possible to resolve objects in biological samples with a size below the diffraction limit of the conventional light microscope (~200 nm). From the perspective of a cell biologist, this is a critical breakthrough; proteins, protein complexes, vesicles and othr small organelles are all in the sub-200 nm size regime. Thus, super-resolution optical microcopy now makes it possible to directly observe these fine sub-cellular features and precisely localize even single molecules in the context of intact fixed and live cells. The OMX Blaze V4 system from Applied Precision enables users to take advantage of two super-resolution imaging modes: structured illumination microscopy (SIM) for fast simultaneous multi- color super-resolution imaging of live cells (lateral resolution = 80-100 nm), and localization microscopy (LM) which provides the highest lateral resolution possible for the imaging of fixed samples (lateral resolution = 20- 50 nm). Major Users (80% usage) are members of the newly formed Epithelial Biology Center (EBC, co- directed by Bob Coffey, M.D. and Jim Goldenring, M.D. Ph.D.); these investigators will apply super-resolution imaging to break new ground in their NIH-funded research projects, which span a wide range of problems in cell and developmental biology, epithelial biology, cancer biology and microbiology. Projects include studies of cytoskeletal dynamics (Kaverina, Ohi, Tyska), polarized membrane trafficking (Coffey, Crowe, Goldenring, Kaverina), mitotic spindle assembly and function (Ohi), assembly of cell surface features such as microvilli and cilia (Goldenring, Tyska), and host-pathogen interactions (Crowe, Tyska). Acquisition of this system will also benefit a group of Minor Users (20% usage) investigating complementary problems at the subcellular level (Janetopoulos, Kenworthy, Miller, Prince, Webb, and Weaver). Together, these investigations hold significance for understanding disease and illness related to birth defects, cancer, bacterial/viral infections, and many other human health problems. The proposed OMX microscope will be installed in the EBC and managed by the Vanderbilt Cell Imaging Shared Resource (CISR, managed by Sam Wells, Ph.D.). With the help of substantial support from the Vanderbilt Office of Research, the CISR will provide the infrastructure for introducing researchers across Vanderbilt (~275 laboratories) to this exciting new technology, training users in the proper operation of this instrument, and applying an established business model to support the long-term maintenance of this system. In summary, acquisition of the OMX will provide unprecedented cellular imaging capabilities that currently do not exist on the Vanderbilt campus. This instrument will enable investigators to image cells with a spatial resolution approaching molecular dimensions, pushing research of biological problems in directions that are simply not possible with the microscopes currently available to Vanderbilt users.