Super-Resolution Structured Illumination Microscope (SIM)

描述（由申请人提供）：细胞生物学与观察细胞器，囊泡，细胞骨架甚至细胞内单个分子的能力密切相关，无论是在正常还是患病状态下。让生物学家看得更清楚的仪器，如共聚焦显微镜和电子显微镜，导致了细胞功能的重大范式转变。最近出现了一种新的成像革命，称为“超分辨率显微镜”，它有能力打破阿贝的“光屏障”，即远场光学显微镜可以解决的问题。这里的潜力是巨大的（类似于20年前的共聚焦显微镜），并有能力解决活细胞中的物体（或固定）与所有的传统荧光标记和GFP探针的彩虹的力量。 我们的用户联盟来自细胞生物学、内分泌学、神经生物学和遗传学，他们都是各自领域的专家，他们要求资金购买蔡司最近商业化的最先进的超分辨率结构照明显微镜（SIM）。初步数据表明，SIM系统将在X，Y和Z轴上的分辨率增加一倍，并提供比传统共焦显微镜高出近8倍的体积分辨率。我们的应用程序非常适合利用这种增强的分辨率，以获得对细胞在天然状态以及糖尿病，癌症和神经病理学中如何发挥作用的新见解。我们联盟的项目集中在三个研究领域：1）神经元，突触和生长锥的成像; 2）膜和膜交通的成像; 3）细胞骨架的成像。我们的目标是使SIM提供给耶鲁大学医学院和更大的科学界的一大群用户。 该系统将在高端定制空间“CINEMA”中心（使用新显微镜方法的细胞成像）的指导下建立，Toomre博士是新型显微镜细胞生物学应用的专家。它将由实验室中训练有素的研究助理（50%的努力）进行适当维护，并由Bewersdorf博士的实验室提供额外的备份，该实验室专门从事超分辨率显微镜的开发和应用。 所要求的仪器预计将是美国同类仪器中的第一台。我们相信这台仪器将为细胞生物学开辟新的途径，并刺激超分辨率显微镜的建立，使其成为生物医学研究的新标准。

DESCRIPTION (provided by applicant): Cell Biology is intimately linked with the ability to see organelles, vesicles, cytoskeleton and even single molecules inside cells, both in normal and diseased states. Instruments that allow biologists to see better, such as the confocal and electron microscope have lead to major paradigm shifts of how cells function. Recently a new revolution in imaging has emerged, called 'super- resolution microscopy' which has the power to break the Abbe's 'light barrier' of what can be resolved by far field light microscopy. The potential here is enormous (akin to confocal microscopes two decades ago) and has the power to resolve objects in living cells (or fixed) with all the power of conventional fluorescent tags and the rainbow of GFP probes. Our consortium of users from cell biology, endocrinology, neurobiology and genetics, whom are experts in their respective fields, request funds to purchase a state-of-the-art super-resolution structured illumination microscope (SIM) that was very recently commercialized by Zeiss. Preliminary data indicate that the SIM system will double the resolution in X, Y, and Z axis and give almost 8-fold better volumetric resolution than conventional confocal microscopes. Our applications are well-matched to capitalize on this enhanced resolution so as to gain new insight into how cells function in both their native state and in diabetes, cancer and neuropathologies. Our consortium's projects focus on three research areas: 1) imaging of neurons, synapses and growth cones; 2) imaging of membranes and membrane traffic; 3) imaging of the cell cytoskeleton. Our aim is to make SIM available to a large group of users at Yale School of Medicine and the larger scientific community. The system will be set up in high-end custom space the 'CINEMA' center (Cellular Imaging using New Microscopy Approaches) under the guidance of Dr. Toomre, an expert in the cell- biological application of novel microscopes. It will be properly maintained by a well-trained Research Associate in the lab (50% effort) with additional backup provided by Dr. Bewersdorf's lab which specializes on super-resolution microscopy development and application. The requested instrument is expected to be the first of its kind in the USA. We believe this instrument will open up new avenues in cell biology and stimulate the establishment of super- resolution microscopy as a new standard in biomedical research.