MRI Development of a fast P3D-STED Microscope

标题：快速P3D-STED显微镜的MRI开发美国国家科学基金会MRI开发补助金授予科罗拉多大学丹佛分校的物理学家、工程师和神经科学家组成的合作团队，以进一步开发受激发射损耗（STED）显微镜的能力。该项目的目标是开发一种FAST P3D-STED显微镜（快速采集，光激活，三维增强分辨率STED显微镜）。从历史上看，光学显微镜的分辨率一直受到衍射势垒的限制，这是一个基本的物理限制，因为光子在通过边缘时会扩散，比如显微镜的孔径。在过去的十年中，出现了几种可以“打破”衍射障壁的方法，在生物成像领域引发了一场革命，并为迄今为止不可能的研究打开了一个重要的机会之窗。Stefan Hell发明的受激发射损耗显微镜（STED）就是这样一种超分辨率技术，可以在几十纳米的分辨率下进行荧光成像。随着技术的不断改进，STED现在能够与广泛的荧光染料和遗传编码荧光团一起使用，并且具有快速，活细胞和活体成像的优势。研究人员将进一步推动STED显微镜的现有能力，开发一种新的STED显微镜，允许高速，亚衍射极限成像和光激活能力相结合。这台显微镜不仅可以让研究人员以亚衍射分辨率观察小物体，还可以实现动态过程的控制和实时观察。该仪器支持的项目包括研究神经可塑性控制下突触中蛋白质复合物的动态组织，通过直接刺激转导途径研究气味转导的分子水平机制，以及在高密度数据存储材料的亚衍射维度上读写比特的方法。这项研究的潜在社会影响包括改善神经系统疾病的治疗和信息处理和存储的进步。FAST P3D STED显微镜将被安置在科罗拉多大学丹佛分校的先进光学显微镜核心（ALMC）设施中，这是一个成功的共享用户设施，拥有人员和资源来培训新用户并操作和维护仪器。科罗拉多大学丹佛分校、科罗拉多大学博尔德分校和其他大学的研究人员、博士后学员和各种科学和工程部门的学生将能够利用显微镜进行研究。与新用户的接触将通过ALMC举办的研讨会和科学会议上的研究报告来完成。研究人员特别致力于促进学生培训和促进少数民族在科学领域的参与，他们参与了神经科学研究成果建设（大脑）项目，这是一个旨在帮助落基山脉和西南地区的不同本科生成功进入神经科学博士项目，从而缩小神经科学研究参与差距的暑期项目。STED显微镜以前所未有的分辨率观察细胞内部运作的能力激发了这些未来科学家的想象力。

Title: MRI Development of a FAST P3D-STED MicroscopeAn NSF MRI Development Grant award is made to a collaborative team of physicists, engineers, and neuroscientists at the University of Colorado Denver to further develop the capabilities of Stimulated Emission Depletion (STED) microscopy. The goal of the project is to develop a FAST P3D-STED microscope (fast acquisition, photoactivation, 3-dimensional enhanced resolution STED microscope). Historically, light microscopy has been limited in its resolution by the Diffraction Barrier - a fundamental physical limit due to the property of photons of spreading out when they pass by an edge, such as the aperture in a microscope. In the past decade, several methods have emerged that can "break" the diffraction barrier, causing a revolution in the field of biological imaging and opening a major window of opportunity for studies that heretofore have been impossible. STimulated Emission Depletion (STED) microscopy, invented by Stefan Hell, is one such super-resolution technique, allowing fluorescence imaging at resolutions of tens of nanometers. With continuing improvements in the technology, STED is now able to be utilized with a wide range of fluorescent dyes and genetically encoded fluorophores and has advantages for fast, live cell, and in vivo imaging. The investigators will further push the current capabilities of STED microscopy by developing a new STED microscope to allow high speed, sub-diffraction limit imaging combined with photoactivation capabilities. This microscope will allow researchers to not only observe small objects with sub-diffraction resolution, but also enable control of dynamic processes and observation in real time.The projects enabled by the instrument include studies of the dynamic organization of protein complexes in synapses upon control of neural plasticity, studies of the molecular-level mechanisms of odor transduction by direct stimulation of the transduction pathway, and methods for writing/reading bits at sub-diffraction dimensions in high density data storage materials. The potential societal impacts of this research include improved treatment of neurological disorders and advances in information handling and storage. The FAST P3D STED microscope will be housed in the Advanced Light Microscopy Core (ALMC) facility at the University of Colorado Denver, a successful shared user facility that has personnel and resources to train new users and operate and maintain the instrument. Researchers, postdoctoral trainees, and students from a variety of science and engineering departments at the University of Colorado Denver, University of Colorado Boulder and other universities will be able to utilize the microscope for their research. Outreach to new users will be accomplished through seminars put on by the ALMC and research presentations at scientific meetings. The investigators are particularly committed to promoting student training and advancing participation of under-represented minorities in science as evidenced by their involvement in the Building Research Achievement in Neuroscience (BRAiN) program, a summer program designed to help meet the challenge to reduce the Neuroscience research participation gap by preparing diverse undergraduates in the Rocky Mountain and Southwest Region for successful entry to Neuroscience Ph.D. programs. The capability of the STED microscope to see the inner workings of cells at unprecedented resolution captures the imagination of these future scientists.