Cell Biology Core

项目总结/摘要 细胞生物学/形态学核心为BADERC研究人员提供技术支持，设备，建议 以及与细胞/膜相关的细胞生物学、免疫细胞化学和形态学研究的专业知识 细胞免疫学和胰岛细胞生物学;血管和内皮细胞 生物学核心位于MGH计划膜生物学设施内，位于Simches 8楼 研究中心，毗邻MGH主校区。核心主任，布朗博士，是一个专家， 膜生物学、膜蛋白和细胞内运输领域。他有一个长期的和有据可查的 过去25年来，MGH核心设施的运营记录，包括20多年来的BADERC核心设施 年该核心的目标是提供对集成细胞成像解决方案和相关专业知识的访问， 糖尿病和其他BADREC目标研究领域的问题，很少或没有孤立的实验室， 资源单独开发。因此，细胞生物学核心旨在最大限度地利用和 专用设备的可用性，这不仅对许多实验室来说过于昂贵， 而且还需要昂贵的定期维护以及使用适当的技术专长。核心 执行从常规免疫荧光成像，共聚焦显微镜，和 （免疫）电子显微镜。核心特别自豪的是，它成功地吸引了NIH共享 多年来的仪器赠款和机构支持。在核心用户的支持下， 在12年多一点的时间里获得了6个SIG，其中最近的一次是在2016年获得了45万美元的新奖项， 蔡司LSM 800 Airyscan共焦平台提供高灵敏度/高分辨率成像和分析 BADERC用户的能力。我们的另一个主要的共焦平台，尼康A1 R共焦系统，提供真实的时间 在温控室中成像、荧光共振能量转移（FRET）和荧光 光漂白后恢复（FRAP）和SIG（30万美元），于2021年5月提交，要求主要 升级得到了20分的优秀分数，应该得到资助。我们还提供激光切割显微切割 和全内反射成像。我们还提供常规的嵌入和切片，以及基本的 免疫荧光孵育专家。核心方案的一个主要和重要目标是提供培训， 来自参与实验室的关键人员，这一策略在之前的资助中非常成功 时期布朗博士和他的高素质的技术支持人员有完善的和行之有效的程序 在不同的项目和特定的实验要求的背景下，最大限度地利用核心。我们特别 注意图像采集和分析的质量控制程序，我们有抗体的协议 验证是否符合当前的严格性和重现性标准。底线是，这个核心继续 像过去一样顺利地工作。它的使用导致了数百个出版物在这段时间内， 由核心工作人员或经过培训的用户拍摄的图像，包括BADERC上一个资助期内拍摄的90多张图像。我们 预计这将继续下去，并根据本申请中提出的用途增加。

PROJECT SUMMARY/ABSTRACT The Cell Biology/Morphology Core provides BADERC investigators with technical support, equipment, advice and expertise for cell biological, immunocytochemical and morphological studies related to cell/membrane biology and signal transduction; cellular immunology and islet cell biology; and vascular and endothelial cell biology. The Core is located within the MGH Program in Membrane Biology facility on the 8th floor of the Simches Research Center, which is adjacent to the MGH main campus. The Core Director, Dr. Brown, is an expert in the area of membrane biology, membrane proteins, and intracellular trafficking. He has a long and well-documented track record of operating Core facilities at MGH over the past 25 years, including this BADERC Core for over 20 years. The goal of this Core is to provide access to integrated cellular imaging solutions and related expertize for problems in Diabetes and other BADERC-targeted research areas that few or no isolated laboratories have the resources to develop individually. The Cell Biology Core is designed, therefore, to maximize the use and availability of specialized pieces of equipment, that are not only prohibitively expensive for many laboratories, but that also require expensive and regular maintenance, and appropriate technical expertise to use. The Core performs all aspects of microscopy from routine immunofluorescence imaging, confocal microscopy, and (immuno)electron microscopy. The Core is especially proud of its record in successfully attracting NIH Shared Instrumentation Grants (SIGs) and institutional support over the years. With support of Core users, we have obtained 6 SIGs in little more than 12 years, the most recent of which was a $450K award in 2016 for a new Zeiss LSM800 Airyscan confocal platform that offers high sensitivity/high resolution imaging and analysis capabilities to BADERC users. Our other major confocal platform, a Nikon A1R confocal system, offers real time imaging in a temperature controlled chamber, fluorescence resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP) and a SIG (for $300K) that was submitted in May 2021 to request major upgrades received an outstanding score of 20 and should be funded. We also offer laser cut microdissection and total internal reflection imaging. We also offer routine embedding and sectioning, and basic immunofluorescence incubation expertize. A major and important objective of the Core is to provide training of key personnel from participating laboratories, a strategy that has worked very successfully over the prior funding periods. Dr. Brown and his highly-qualified technical support staff have well-established and proven procedures for maximizing Core use in the context of varied projects and specific experimental requirements. We pay special attention to quality control procedures for image capture and analysis, and we have protocols for antibody verification to comply with current Rigor and Reproducibility standards. The bottom line is that this Core continues to work smoothly, as in the past. Its use has resulted in several hundred publications over that time that contain images taken by Core staff, or by trained users, including over 90 in the previous BADERC funding period. We expect this to continue and increase based on the usage proposed in this application.