HD Upgrade to a Nikon A1R Confocal Imaging Platform

高清升级至尼康 A1R 共焦成像平台

• 批准号: 10415591
• 负责人: Dennis Brown
• 金额: $ 29.61万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10415591
• 关键词: 10 year old; 3-Dimensional; Applications Grants; Cells; Community Hospitals; Confocal Microscopy; Core Facility; Coupled; Exposure to; Funding; General Hospitals; Health; Image; Imaging Techniques; Individual; Lasers; Leadership; Massachusetts; Membrane Biology; Microscope; Microscopy; Modernization; Peer Review; Photobleaching; Proteins; Publications; Research; Resolution; Savings; Scanning; Specimen; Speed; System; Techniques; Time; United States National Institutes of Health; base; cellular imaging; confocal imaging; detector; experimental study; image processing; imaging platform; improved; instrument; live cell imaging; multidisciplinary; programs; ranpirnase; reconstruction; solid state; time use

Project Summary We request funds to support the upgrade of our existing Nikon A1R confocal microscope, which is now almost 10 years old and lacks many of the newer features that have been developed since its purchase in 2012. The upgraded instrument will be housed within the Program in Membrane Biology (PMB) Microscopy Core at the Massachusetts General Hospital (MGH) in the multidisciplinary Simches Research Center building. The mandate of this MGH-approved Core facility is to provide access to a full range of microscopy techniques to 2 NIH funded P30 Centers and individual research groups within the MGH community and beyond. The Core has considerable expertise in static and live cell imaging techniques used by our extensive user-base over the years. Our imaging capabilites were enhanced 4 years ago by the acquisition of a Zeiss LSM800 Airyscan confocal platform to function alongside the older Nikon A1R. Together these instruments have provided sufficient high level imaging bandwidth to serve our users. The Nikon A1R is extensively used for longer term, live cell imaging and has an inverted microscope and an environmental chamber to faciliate this need. The LSM800, associated with an upright microscope and no environmental chamber, provides consistent and timely access to a high resolution confocal that is mostly used to perform static imaging of fixed specimens. With the increase in demand for imaging of living cells expressing various types of fluorescent molecules and tagged proteins, and the increasing need to image these cells over time using 3D reconstructions, the purpose of this application is to provide our users with access to modern confocal microscopy options that are better adapted for specialized imaging purposes than the current older A1R system. In particular, the new high speed HD resonant scanning mode with associated AI image processing capabilities provides a remarkable increase in image size and resolution when compafed to the old resonant scanning system. Coupled with replacement of the old PMT detectors with new GaAsp detectors and the incorporation of stable solid state lasers, these upgrades will allow specimens to be exposed to much lower laser intensities, reducing photobleaching and photodamage, while providing a significant increase in resolution compared to the old system. In addition, images and multicolor 3D stacks can be collected much more rapidly in resonant scanning mode, while not sacrificing quality, meaning that experiments can be conducted quickly and efficiently, saving time and money for our users. The PMB Core has supported hundreds of peer-reviewed publications and dozens of grant applications over the years. The Core has an ongoing quest to constantly improve and upgrade Core facilities in support of its user base. This has been achieved using funds from NIH SIG applications as well as considerable institutional support from MGH leadership. Providing our Core users with continued access to up-to-date confocal imaging is an essential part of maintaining the MGH community at the cutting edge of NIH-funded, health-related research.

项目摘要 我们要求资金支持我们现有的尼康A1R共聚焦显微镜的升级，这是现在 几乎10岁，缺乏许多新的功能，已被开发，因为它的购买， 2012.升级后的仪器将被安置在膜生物学（PMB）显微镜计划内 多学科Simches研究中心的马萨诸塞州总医院（MGH）核心 建设该MGH批准的核心设施的任务是提供全方位的显微镜检查 技术2 NIH资助的P30中心和MGH社区内的个别研究小组， 超越。核心在静态和活细胞成像技术方面具有相当的专业知识， 多年来，广泛的用户基础。我们的成像能力在4年前通过收购 蔡司LSM800 Airyscan共焦平台与老款尼康A1R一起使用。综合这些 仪器提供了足够的高水平成像带宽，以服务于我们的用户。尼康A1R 广泛用于长期活细胞成像，并具有倒置显微镜和环境 会议室，以满足这一需求。LSM 800与直立显微镜相结合， 室，提供一致和及时的访问高分辨率共焦，主要用于执行 固定标本的静态成像。随着对表达各种细胞的活细胞成像需求的增加， 荧光分子和标记蛋白的类型，以及随着时间的推移对这些细胞成像的需求越来越大 使用3D重建，这个应用程序的目的是为我们的用户提供现代的访问 共聚焦显微镜的选择，更好地适应专门的成像目的比目前的旧 A1R系统。特别是，新的高速HD共振扫描模式与相关的AI图像 处理能力提供了显着增加的图像大小和分辨率时，相比， 旧的共振扫描系统再加上用新的GaAsp取代旧的PMT探测器 探测器和稳定的固态激光器的结合，这些升级将使标本 暴露于低得多的激光强度，减少光漂白和光损伤，同时提供 与旧系统相比，分辨率大幅提高。此外，图像和3D堆栈 可以在谐振扫描模式下更快地收集，同时不牺牲质量，这意味着， 实验可以快速有效地进行，为我们的用户节省时间和金钱。PMB核心 多年来，它支持了数百篇同行评审的出版物和数十项资助申请。的 核心一直在寻求不断改进和升级核心设施，以支持其用户群。 这是利用NIH SIG申请的资金以及相当大的机构支持实现的 MGH的领导。为我们的核心用户提供持续访问最新的共焦成像是一个 保持MGH社区在NIH资助的，健康相关研究的前沿的重要组成部分。