ORD Shared Equipment Evaluation Program (ShEEP) (IS1) - Zeiss LSM980 Airyscan Confocal Microscope

退伍军人事务部田纳西谷医疗保健系统(VATVHS)，纳什维尔校区正在申请资金购买 蔡司LSM980空气扫描共焦显微镜。LSM980是一款最先进的点扫描共焦 具有超分辨率、活细胞和大型马赛克成像的特殊功能的显微镜。LSM980 将可供退伍军人管理局调查人员使用，并由范德比尔特细胞成像共享资源(CISR)维护。 这一新仪器的增加对支持八名退伍军人管理局调查人员正在进行的研究至关重要 在此建议书中表示为主要用户项目。这些调查人员都有活跃的退伍军人管理局资金 基本需要推进研究课题，包括糖尿病、艰难梭菌感染、炎症性肠炎 疾病(IBD)、细胞外基质和纤维化、慢性阻塞性肺疾病(COPD)、肾脏 发展，和胃癌。我们预计退伍军人管理局和美国政府内部的许多其他研究项目 更广泛的范德比尔特社区也将从这一工具中受益；特别是那些需要超级- 分辨率、活样本和大组织成像。这款蔡司LSM980 AiryScan将提供专用共焦 这些退伍军人研究人员的工作时间以及AiryScan提供的更高分辨率和更高的超级 分辨率成像速度。我们选择LSM980作为这8个退伍军人管理局用户的重要需求，基于以下三点 主要目标：1)提供额外的共焦时间，因为现有的CISR共焦显微镜已饱和， 2)增加一个共焦显微镜，专为退伍军人管理局的调查人员提供，他们需要更长的成像时间 样品和活细胞成像，3)提供新功能，包括最新一代的4x AiryScan 更快的超分辨率成像和最先进的探测器，提高了灵敏度。所请求的 仪器是根据主要用户的需求配置的，咨询了PI、Roy Zent博士和CELL 正在为共享资源人员创建映像。完整的系统包括LSM980共焦与最先进的 GaAsP PMTS、AiryScan超分辨率模块、孵化和环境控制、各种目标 镜头、6台激励激光器和一个装有Zen 3.1软件的采集工作站。重要注意事项 在选择各种物镜以容纳样品时，考虑到了光学性能 包括固定的或活的细胞、固定的或活的组织和大的光学透明组织。最新一代的 探测器为信号检测提供更高的灵敏度，并为活细胞实验提供温和的成像。新的 Airyscan2模块提供单独的探测器，分辨率最高可达1.7倍 常规共聚焦显微镜。环境舱配备37℃孵化，5%二氧化碳，以及 热漂移校正允许实时细胞成像，从几分钟到几个小时到一夜。激励激光器包括 405、445、488、514、561和639 nm适合广泛的现代荧光蛋白质和 染料。这些LSM980配件是成功实施先进成像的基础 在主要用户项目中进行了描述。对这一工具的专业知识和专门支持是特别的。 该研究所已有20多年的历史，为400多个拥有尖端成像专业知识的调查实验室提供协助 和合作研究支持。Pi Roy Zent博士是一名内科医生和退伍军人管理局调查员，具有以下专业知识 肾脏发育和高分辨率显微镜，是该项目的主要用户。此外，所有8大调 该方案中的用户是具有共焦显微镜经验的退伍军人管理局调查人员。

The VA Tennessee Valley Healthcare System (VATVHS), Nashville campus is requesting funds to purchase a Zeiss LSM980 Airyscan confocal microscope. The LSM980 is a state-of-the-art point-scanning confocal microscope with particular functionality for super-resolution, live cell, and large mosaic imaging. The LSM980 will be available for VA investigators and maintained by the Vanderbilt Cell Imaging Shared Resource (CISR). The addition of this new instrument is crucial to support the ongoing research of eight VA investigators represented as Major Users Projects in this proposal. These investigators all have active VA funding and essential needs to advance research topics including diabetes, C. difficile infection, Inflammatory Bowel Disease (IBD), extracellular matrix and fibrosis, chronic obstructive pulmonary disease (COPD), kidney development, and gastric cancer. We expect that numerous other research programs at the VA and within the wider Vanderbilt community will also benefit from this instrument; particularly those that require super- resolution, live sample, and large tissue imaging. This Zeiss LSM980 Airyscan will offer dedicated confocal hours for these VA researchers as well as greater resolution afforded by Airyscan and increased super- resolution imaging speeds. We chose the LSM980 as a significant need for these 8 VA users based on three main goals: 1) to provide additional confocal hours because existing CISR confocal microscopes are saturated, 2) to add a confocal microscope dedicated to VA investigators who need longer imaging sessions for large samples and for live cell imaging, 3) to offer new capabilities including the latest generation Airyscan for 4x faster super-resolution imaging and state-of-the-art detectors for increased sensitivity. The requested instrument is configured based on the needs of the Major Users in consultation with PI Dr. Roy Zent and Cell Imaging Shared Resource personnel. The full system includes the LSM980 confocal with state-of-the-art GaAsP PMTs, Airyscan super-resolution module, incubation and environmental control, a variety of objective lenses, 6 excitation lasers, and an acquisition workstation with ZEN 3.1 software. Significant consideration was given to optical performance in choosing the variety of objective lenses to accommodate samples including fixed or live cells, fixed or live tissue, and large optically cleared tissue. The latest generation of detectors offers greater sensitivity for signal detection and gentle imaging for live cell experiments. The new Airyscan2 module offers a separate detector to achieve resolution up to 1.7x greater than the limit of conventional confocal microscopy. The environmental chamber equipped with 37 C incubation, 5% CO2, and thermal drift correction allows live cell imaging from minutes to hours to overnight. Excitation lasers including 405, 445, 488, 514, 561, and 639 nm are appropriate for a wide range of modern fluorescent proteins and dyes. These LSM980 accessories are fundamental to the successful implementation of the advanced imaging described in the Major User projects. The expertise and dedicated support for this instrument are exceptional. The CISR has a 20+ year history of assisting over 400 investigator labs with cutting-edge imaging expertise and cooperative research support. PI Dr. Roy Zent is a Staff Physician and VA investigator with expertise in kidney development and high-resolution microscopy and is Major User on this project. Additionally, all 8 Major Users in this proposal are VA investigators with confocal microscopy experience.