Super-Resolution Structured Illumination Microscope (SIM)

描述（由申请人提供）：细胞生物学与观察细胞内的细胞器、囊泡、细胞骨架甚至单个分子的能力密切相关，无论是在正常状态还是病变状态。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，已经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“光障”，可以通过远场光显微镜解决。这里的潜力是巨大的（类似于二十年前的共聚焦显微镜），并且具有传统荧光标签和彩虹绿色荧光蛋白探针的所有功能，可以分辨活细胞（或固定细胞）中的物体。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，他们是各自领域的专家，要求资金购买最先进的超分辨率结构照明显微镜（SIM），该显微镜最近由蔡司商业化。初步数据表明，SIM系统在X， Y和Z轴上的分辨率将提高一倍，并且比传统共聚焦显微镜的体积分辨率提高近8倍。我们的应用可以很好地利用这种增强的分辨率，从而获得新的见解，了解细胞在其天然状态下以及在糖尿病、癌症和神经病理中的功能。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)膜成像与膜交通；3)细胞骨架成像。我们的目标是将SIM提供给耶鲁医学院和更大的科学界的一大群用户。该系统将在新型显微镜的细胞生物学应用专家Toomre博士的指导下，在高端定制空间“CINEMA”中心（使用新显微镜方法的细胞成像）建立。它将由一名训练有素的研究助理在实验室（50%的努力）进行适当的维护，Bewersdorf博士的实验室专门从事超分辨率显微镜的开发和应用，提供额外的支持。所要求的仪器预计将是美国同类仪器中的第一个。我们相信该仪器将开辟细胞生物学的新途径，并促进超分辨率显微镜作为生物医学研究的新标准的建立。

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.