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

弗吉尼亚州田纳西山谷医疗保健系统（VATVHS），纳什维尔校园正在申请资金购买一个 Zeiss LSM980 Airyscan共聚焦显微镜。LSM 980是一款最先进的点扫描共焦 显微镜具有超分辨率、活细胞和大型马赛克成像的特殊功能。LSM980 将提供给VA研究者，并由范德比尔特细胞成像共享资源（CISR）维护。 这一新仪器的加入对于支持八名VA研究人员正在进行的研究至关重要 在本建议书中被称为主要用户项目。这些调查人员都有积极的VA资金， 基本需要推进研究课题，包括糖尿病，C。艰难梭菌感染，肠道炎症 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，许多其他研究计划在弗吉尼亚州和内部 更广泛的范德比尔特社区也将受益于这一工具，特别是那些需要超级， 分辨率、活体样本和大组织成像。这款蔡司LSM980 Airyscan将提供专用的共焦 小时，以及Airyscan提供的更高分辨率和更高的超分辨率， 分辨率成像速度。我们选择LSM 980作为这8个VA用户的重要需求，是基于以下三点： 主要目标：1）由于现有的CISR共焦显微镜已经饱和， 2)增加一个共聚焦显微镜，专门用于VA调查人员，他们需要更长的成像时间， 3）提供新功能，包括最新一代Airyscan for 4x 更快的超分辨率成像和最先进的探测器，以提高灵敏度。请求的 根据主要用户的需求配置仪器，并与PI Dr. Roy Zent和Cell协商 成像共享资源人员。完整的系统包括LSM 980共焦， GaAsP光电倍增管，Airyscan超分辨率模块，孵育和环境控制，多种物镜 镜头，6个激发激光器和一个带有ZEN 3.1软件的采集工作站。重要考虑 在选择各种物镜以适应样品时， 包括固定的或活的细胞、固定的或活的组织以及大的光学透明组织。最新一代 探测器为信号检测提供了更高的灵敏度，并为活细胞实验提供了温和的成像。新 Airyscan2模块提供了一个单独的探测器，可实现高达1.7倍的分辨率， 传统的共聚焦显微镜。环境室配备有37 ℃孵育，5%CO2， 热漂移校正允许活细胞成像从数分钟到数小时到过夜。激发激光器，包括 405、445、488、514、561和639 nm适用于广泛的现代荧光蛋白， 染料。这些LSM 980附件是成功实施高级成像的基础 在主要用户项目中描述。对这一文书的专门知识和专门支持是非同寻常的。 CISR拥有20多年的历史，为400多个研究实验室提供尖端的成像专业知识 合作研究支持。PI Roy Zent博士是一名工作人员医生和VA研究员，擅长于 肾脏发育和高分辨率显微镜，是这个项目的主要用户。此外，8大 本提案中的用户是具有共聚焦显微镜经验的VA研究人员。

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.