IDBR: Development of a High Resolution Serial Block-Face Scanning Electron Microscope

IDBR：高分辨率串行块面扫描电子显微镜的开发

• 批准号: 0964114
• 负责人: Eduardo Rosa-Molinar
• 金额: $ 45万
• 依托单位: University of Puerto Rico-Rio Piedras
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0964114&HistoricalAwards=false
• 关键词: IDBR; Development; Resolution; Serial; Block

Detailed circuit maps of neural networks and their synaptic connections between/among neurons of the central nervous system (CNS) are needed to advance CNS studies. This need is particularly acute in the emerging field of "connectomics" that requires a very large number of high-resolution three-dimensional microscopy wide-field images to visualize and analyze CNS neural circuitry. Instruments such as Focused Ion Beam Serial Block Face Scanning Electron Microscopy (FIB-SBFSEM) and serial section transmission electron microscopy (ssTEM) achieve high-resolution three-dimensional images and provide the resolution required to trace fine dendrites and synapses. However, Serial Block Face Scanning Electron Microscopy (SBFSEM) is the only three-dimensional microscopy technique that can provide wide-field images. A single instrument that provides high-resolution three-dimensional wide-field images is acutely needed. The principal investigator (PI) in concert with JEOL Inc. will take the first crucial step to meet that need by improving a thru-the-lens (TTL) backscatter detector that will increase the backscattered electron image (BEI) resolution and increase image acquisition speed. Intellectual merit: Improvements in the TTL backscatter detector will greatly advance neuroscientists' ability to 1) identify specific spinal neuron phenotypes and 2) gain insights into the 'impenetrable jungles' of central nervous system regions, thus overcoming obstacles identified over 100 years ago. Broader Impacts: TTL backscatter detector improvements will: 1) permit mapping of basic synaptic microcircuits of some regions of the central nervous system; 2) be used to train next generation Hispanic scientists in the emerging field of connectomics; 3) meet 30 Puerto Rican neuroscientists' need for access to instrumentation; upon successful completion of the beta test and with development and marketing, the instrument will 4) meet the need of many, many neuroscientists engaged in connectivity and other CNS studies. Dissemination: The PI will offer Hispanic neurobiology graduate students and 30 neurobiology faculty members training on the new instrument. The PI's laboratory provides its research images to industry which uses them in its brochures, appropriately crediting the PI and funding agents. This arrangement will continue and expand. The TTL backscatter detector will create opportunities to exploit stunning visual images as a way of enhancing awareness of new developments in science. Towards this goal we will: 1) use images generated from these projects as artwork in UPR-RP class and conference rooms; 2) submit articles to the UPR-RP alumni magazine and to Inventio, a bi-lingual campus publication that presents research and scholarly achievements to the larger business and industry community and to the general public; and 3) use the images to persuade local newspapers and magazines to do more stories on basic science. We hope to use riveting and beautiful images to capture the imagination and interest of the lay public. Finally, through a collaboration with the mouse Biomedical Imaging Research Network (mouse BIRN) in the Laboratory of Neuro Imaging (LONI) at University of California Los Angeles (UCLA), we use the Mouse BIRN Atlasing Toolkit (MBAT), a platform independent image viewer with the ability to simultaneously present multiple images from multiples sources on a single viewing "canvas". With MBAT, local and online images can be compared and explored side-by-side. MBAT allows almost anyone with a modest personal computer and internet connection to view virtual sections of our image data on our website (http://pisces.cnnet.clu.edu-lab/final/).

详细的神经网络电路图和它们在中枢神经系统（CNS）的神经元之间的突触连接是推进CNS研究所必需的。 这种需求在新兴的“连接组学”领域尤其迫切，该领域需要大量的高分辨率三维显微镜宽视场图像来可视化和分析CNS神经回路。 聚焦离子束连续块面扫描电子显微镜（FIB-SBFSEM）和连续切片透射电子显微镜（ssTEM）等仪器可获得高分辨率三维图像，并提供追踪精细树突和突触所需的分辨率。 然而，连续块面扫描电子显微镜（SBFSEM）是唯一的三维显微镜技术，可以提供宽场图像。 迫切需要一种能够提供高分辨率三维宽视场图像的单一仪器。 主要研究者（PI）与JEOL Inc.将采取第一个关键步骤，以满足这一需要，通过改善通过透镜（TTL）的反向散射探测器，将增加反向散射电子图像（BEI）的分辨率和提高图像采集速度。 智力优点：TTL反向散射探测器的改进将大大提高神经科学家的能力，1）识别特定的脊髓神经元表型，2）深入了解中枢神经系统区域的“不可穿透的丛林”，从而克服100多年前发现的障碍。更广泛的影响：TTL反向散射探测器的改进将：1）允许绘制中枢神经系统某些区域的基本突触微电路; 2）用于在新兴的连接组学领域培训下一代西班牙裔科学家; 3）满足30名波多黎各神经科学家对仪器的需求;在成功完成β测试后，随着开发和销售，该仪器将满足许多从事连接性和其他中枢神经系统研究的神经科学家的需要。传播：PI将为西班牙裔神经生物学研究生和30名神经生物学教员提供新仪器的培训。PI的实验室将其研究图像提供给在其小册子中使用它们的行业，适当地归功于PI和资助代理人。这一安排将继续下去并扩大。 TTL反向散射探测器将创造机会，利用惊人的视觉图像作为提高对科学新发展的认识的一种方式。 为实现这一目标，我们将：1）使用从这些项目产生的图像作为艺术品在UPR-RP类和会议室; 2）提交文章到UPR-RP校友杂志和Inventio，双语校园出版物，介绍研究和学术成果，以更大的商业和工业界和公众;和3）使用图像说服当地报纸和杂志做更多的故事基础科学。 我们希望用引人入胜和美丽的图像来捕捉公众的想象力和兴趣。最后，通过与加州洛杉矶大学（UCLA）神经成像实验室（LONI）的小鼠生物医学成像研究网络（小鼠BIRN）的合作，我们使用了小鼠BIRN浏览工具包（MBAT），一个平台独立的图像查看器，能够同时呈现多个图像从多个来源在一个单一的查看“画布”。通过MBAT，本地图像和在线图像可以并排进行比较和探索。 MBAT允许几乎任何拥有普通个人电脑和互联网连接的人在我们的网站（http：//pisces.cnnet.clu.edu-lab/final/）上查看我们图像数据的虚拟部分。