ACQUISITION OF A ZEISS CONFOCAL MICROSCOPE

在现代细胞生物学研究中，获取、处理、分析和显示固定细胞和活细胞的三维图像的能力已经成为一种普遍而关键的需求。在确定的解剖位置、细胞过程和结构的背景下，对选定分子的外观、位置、运动和周转的详细分析提供了无法通过其他方式获得的基本信息。共聚焦激光扫描显微镜以三维图像堆栈的形式提供这些信息，并且可以同时对多个不同的荧光信号进行成像。我们的机构目前还没有共聚焦显微镜，并且该提案所代表的研究人员迄今为止依赖于合作或越来越有限地使用单一的本地共聚焦仪器，以推进我们在需要三维成像的领域的研究项目。本提案中概述的具体研究项目涉及秀丽隐杆线虫的神经解剖学发育，培养和生发中心B细胞中抗体产生的表达和成熟，骨髓中B细胞的发育，内皮细胞上和内皮细胞上凝血蛋白的调节和功能，以及导致DT40 B细胞颗粒摄取的信号通路。研究人员对高分辨率的三维成像有直接的、持续的需求，并且在任何情况下都有共聚焦显微镜的经验。所要求的系统将安置在OMRF核心成像设施内，其使用将由训练有素的成像专家监督，以便于维修、培训和使用。这一系统的可用性将对欧姆研究基金正在进行的研究的质量和数量产生重大影响。

The ability to acquire, process, analyze and display three-dimensional images of fixed and of living cells has become a common and critical need in modern cell biological research. Detailed analyses of the appearance, location, movements and turnover of selected molecules in the context of defined anatomical locations, cellular processes and structures provide essential information that cannot otherwise be obtained. Confocal laser- scanning microscopy provides this information in the form of three- dimensional image stacks and can simultaneously image multiple different fluorescent signals. No confocal microscope is currently available at our institution and the investigators represented by this proposal have so far depended on collaborations or increasingly limited access to the single locally-available confocal instrument in order to advance our research programs in areas that require three-dimensional imaging. The specific research projects outlined in this proposal address neuroanatomical development in C. elegans, expression and maturation of antibody production in B cells in culture and in germinal centers, B cell development in bone marrow, regulation and function of blood coagulation proteins on and in endothelial cells, and signaling' pathways leading to particle uptake in DT40 B cells. The investigators have immediate, ongoing needs for three-dimensional imaging at high resolution and in every case have prior experience with confocal microscopy. The requested system will be housed in the OMRF Core Imaging Facility and its use will be supervised by a well-trained Imaging Specialist in order to facilitate maintenance, training and access. The availability of this system will have a great impact on the quality and quantity of ongoing research at the OMRF.