Super-Resolution Structured Illumination Microscope (SIM)

描述(申请人提供)：细胞生物学与观察细胞内细胞器、小泡、细胞骨架甚至单分子的能力密切相关，无论是正常状态还是疾病状态。让生物学家看得更清楚的仪器，如共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，成像领域出现了一场新的革命，称为“超分辨率显微镜”，它有能力打破阿贝的“光障”，这是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年前的共焦显微镜)，并具有解析活细胞中物体的能力(或固定)，具有传统荧光标签和GFP探头的彩虹。我们的用户来自细胞生物学、内分泌学、神经生物学和遗传学，他们都是各自领域的专家，他们申请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系统将把X、Y和Z轴的分辨率提高一倍，并提供比传统共焦显微镜高近8倍的体积分辨率。我们的应用非常适合利用这一增强的分辨率，以便对细胞在其自然状态下以及在糖尿病、癌症和神经病理中的功能有新的了解。我们联盟的项目集中在三个研究领域：1)神经元、突触和生长锥体的成像；2)膜和膜交通的成像；3)细胞细胞骨架的成像。我们的目标是将SIM提供给耶鲁医学院的一大批用户和更大的科学界。该系统将在新型显微镜的细胞生物学应用专家图姆雷博士的指导下，在高端定制空间“电影”中心(使用新的显微镜方法进行细胞成像)中建立。它将由实验室中训练有素的研究助理(50%的努力)进行适当的维护，并由比沃斯多夫博士的实验室提供额外的支持，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第一个此类仪器。我们相信，这台仪器将为细胞生物学开辟新的途径，并推动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.