Acquisition of a supercontinuum laser STED microscope

描述（由申请人提供）：我们申请资金购买超连续光谱显微镜。Stefan Hell发明的STED（受激发射损耗）显微镜是第一台突破阿比衍射极限的光学显微镜。阿比势垒是一条基本的物理定律，它指出，一个无限小的光源，比如一个1纳米的荧光分子，不能被成像成一个比大约300纳米宽小得多的点，因为光子在经过边缘时会扩散。因此，任何尺寸较小的结构都会出现衍射受限的模糊点。由于许多对生物学感兴趣的结构都是在较小的尺度上，这是光学显微镜的一个严重限制。Stefan Hell （Max-Planck Institute for生物物理化学，Gvttingen）用STED显微镜解决了光学衍射极限的问题。通过这项技术，他证明了XY分辨率提高了大约10倍，这一进步为医学和生物科学的研究打开了一扇巨大的机会之窗，这在以前是不可能的。我们相信，我们是唯一适合获得这种新的STED显微镜。我们是一个由12名经验丰富，独立资助的研究人员组成的团队，他们在神经生物学，细胞生物学，分子生物学以及先进的光学技术方面具有专业知识。我们已与S. Hell建立了密切的工作关系，他已同意担任高级顾问。Restrepo （PI）在格廷根与他一起研究了一个发表在《美国科学院院刊》上的项目。此外，Betz还培养了S. Hell的一位重要同事Silvio Rizzoli（2004年美国科罗拉多大学博士），他曾是Hell博士的博士后，现为格廷根欧洲神经科学研究所的组长（相当于助理教授）。里佐利已经同意担任技术顾问。Emily Gibson （Co-PI）和Tim Lei是科罗拉多大学丹佛分校的技术顾问，在非线性激光光学方面都有丰富的经验。吉布森将访问S. Hell在格廷根的实验室，接受关于超连续统标准实施的培训。我们所有的用户都是美国国立卫生研究院资助的研究者。所有项目的重点是基础生物学和疾病相关研究。本申请中列出的每个拟议项目都是高影响力的研究，将推进当前的科学理解。这些项目不能用我们现有的任何其他工具来完成。STED显微镜将被纳入科罗拉多大学丹佛分校现有的核心光学显微镜设施（LMF）。六年来，LMF已经满足了2000多名教职员工和学员的需求，目前有四个先进的荧光显微镜。经验丰富、知识渊博的LMF人员确定时间分配和用户费用，吸引和培训新用户，确保安全，管理账单和友好的在线调度。当安装新仪器时，特殊的启动活动确保其顺利融入LMF。

DESCRIPTION (provided by applicant): We request funds to purchase a supercontinuum STED microscope. The STED (STimulated Emission Depletion) microscope, invented by Stefan Hell, is the first light microscope to break the Abbi diffraction limit. Abbi's barrier, a fundamental law of physics, states that an infinitesimally small source of light, e.g. a single fluorescing 1 nm molecule, cannot be imaged to a spot much smaller than roughly 300 nm wide, because photons spread out as they pass by an edge. Thus, any structure smaller in size will appear as a diffraction- limited blurred spot. Since many structures of biological interest are on a smaller scale, this is a crippling limitation to light microscopy. Stefan Hell (Max-Planck Institute for Biophysical Chemistry, Gvttingen) solved the problem of the optical diffraction limit with the STED microscope. With this technique, he has demonstrated a roughly ten-fold increase in XY resolution, an advance that opens a huge window of opportunity for studies in the medical and biological sciences that heretofore have been impossible. We believe that we are uniquely suited to acquire this new STED microscope. We are a group of 12 experienced, independently-funded researchers with expertise in neurobiology, cell biology, molecular biology, as well as advanced optical technologies. We have established a close working relationship with S. Hell who has agreed to serve as a Senior Consultant. Restrepo (PI) worked with him in Gvttingen on a project published in PNAS. In addition, Betz, a Major User, trained a key colleague of S. Hell's, Silvio Rizzoli (PhD, University of Colorado, 2004), former postdoc with Dr. Hell, now Group Leader (Assistant Professor equivalent) at the European Neuroscience Institute, Gvttingen. Rizzoli has agreed to serve as Technical Consultant. Emily Gibson (Co-PI) and Tim Lei a Technical Consultant at CU Denver are both experienced in nonlinear laser optics. Gibson will visit S. Hell's lab in Gvttingen to be trained on the implementation of the supercontinuum STED. All our Users are NIH-funded investigators. The focus of all projects is in basic biological and disease related research. Each proposed project outlined in this application is high impact research that will advance current scientific understanding. These projects cannot be accomplished with any other instrument currently available to us. The STED microscope will be incorporated into the existing core Light Microscopy Facility (LMF) at the University of Colorado Denver. For six years, the LMF has served the needs of more than 2000 faculty, staff, and trainees, currently with four advanced fluorescence microscopes. Experienced and knowledgeable LMF personnel determine time allocation and user fees, attract and train new users, insure security, and manage billing and a friendly online scheduler. When a new instrument is installed, special start-up activities insure its smooth incorporation into the LMF.