Multi-purpose Superresolution Microscope for Core Facility

描述（由申请人提供）：多年来，显微镜已经得到了改进，可以提供更高的灵敏度和精度，但光学透镜的焦斑衍射极限直到最近才被打破。受激辐射损耗（STED）技术是一种物理结构的照明焦点，以产生有效的照明点小于3-4倍的衍射极限。就像共聚焦显微镜提供的分辨率是标准显微镜的2-3倍一样，STED的“超分辨率”至少比共聚焦显微镜高出2-3倍，并且更接近电子显微镜的分辨率。芝加哥大学的集成光学显微镜设施（ILMF）为160多名美国国立卫生研究院资助的生物科学部研究人员以及来自物理科学部和邻近机构的跨学科研究人员提供服务。在过去的十年里，ILMF见证了共聚焦显微镜使用的爆炸式增长。尤其受欢迎的是ILMF中最先进的仪器：徕卡SP5多光子共聚焦显微镜，它通常被预订一空，必须提前很久预订。本提案中要求的徕卡SP5 II STED-CW显微镜是SP5的增强版，包括更高的灵敏度，最重要的是，提供实时超分辨率能力。仪器

DESCRIPTION (provided by applicant): Microscopes have improved over the years to permit ever greater sensitivity and precision, yet the focal spot diffraction limit of optical lenses has ot been broken until very recently. Stimulated Emission Depletion (STED) technology is a physical structuring of the illumination focus to produce effective illumination points 3-4 times smaller than the diffraction limit. Just as confocal microscopy provides 2-3 times the resolution of standard microscopy, STED "superresolution" outperforms confocal microscopy by at least this margin and closer approaches the resolution of the electron microscope. The Integrated Light Microscopy Facility (ILMF) at The University of Chicago serves more than 160 NIH-funded researchers in the Biological Sciences Division, as well as interdisciplinary researchers from the Physical Sciences Division and neighboring Institutions. During the past decade the ILMF has witnessed an explosive growth in the use of confocal microscopy. Particularly popular is the most advanced instrument in the ILMF: a Leica SP5 multiphoton confocal microscope, which is typically booked to capacity and must be reserved far in advance. The Leica SP5 II STED-CW microscope requested in this proposal is an enhanced version of the SP5 that includes greater sensitivity, and, most significantly, provides real-time superresolution capability. The instrument includes the (newer) continuous-wave depletion laser scheme that is better suited for live-cell probes. The system includes the latest technology, a hybrid detector that offers APD-like sensitivity with lower noise and wider dynamic range. The researchers at the University of Chicago require the resolution enhancement as well as the increased capacity, and have undertaken a lease to obtain these tools while replacing a failing, older SP2 confocal microscope in a Core serving 162 PIs and over 750 users, an action that demonstrates strong Institutional Commitment for this proposal. The seamlessly integrated STED capability is a major technical advance for researchers at The University of Chicago, and is a novel regional resource also available to researchers from other Institutions. PUBLIC HEALTH RELEVANCE: This request is to buyout an interim lease that the University of Chicago has undertaken to provide continued basic and newly-advanced high-end research microscopy, replacing a failing older system. The requested superresolution microscope has undergone extensive in-house testing, is housed in an established Core Facility, and provides novel capabilities that will significantly enhance the research programs of a large NIHfunded user community.