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 我们请求资金支持我们现有的尼康A1R共焦显微镜的升级，这是现在 已有近10年的历史，缺乏许多自 2012年。升级后的仪器将安装在膜生物学(PMB)显微镜计划中 马萨诸塞州综合医院(MGH)多学科西姆斯研究中心的核心 大楼。MGH批准的核心设施的任务是提供对全方位显微镜的访问 为国立卫生研究院资助的2个P30中心和MGH社区内的个人研究小组提供技术 更远一点。酷睿在静态和活细胞成像技术方面拥有丰富的专业知识，我们的 多年来拥有广泛的用户基础。我们的成像能力在4年前因此次收购而得到增强 拥有蔡司LSM800 Airycan共焦平台，可与老式尼康A1R一起工作。把这些放在一起 仪器提供了足够的高水平成像带宽来服务我们的用户。尼康A1R是 广泛用于更长期的活细胞成像，并具有倒置显微镜和环境 为满足这一需要而设立的商会。LSM800，与立式显微镜相关联，不受环境影响 提供一致和及时的高分辨率共焦，主要用于执行 固定样品的静态成像。随着对表达不同种类的活细胞的成像需求的增加 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进行成像的需求日益增加 使用3D重建，此应用程序的目的是为我们的用户提供访问现代 共聚焦显微镜选项，比当前较旧的更适合专门成像目的 A1R系统。特别是，新的高速高清共振扫描模式与关联的AI图像 与传统的 旧的共振扫描系统。加上用新的GaAsp取代旧的PMT探测器 探测器和稳定固态激光器的结合，这些升级将使样品能够 暴露在低得多的激光强度下，减少光漂白和光损伤，同时提供 与旧系统相比，分辨率显著提高。此外，图像和多色3D堆栈 可以在共振扫描模式下更快地收集，而不会牺牲质量，这意味着 实验可以快速高效地进行，为我们的用户节省时间和金钱。港口及航运局的核心 多年来支持了数百份同行评议的出版物和数十份赠款申请。这个 核心不断改进和升级核心设施，以支持其用户基础。 这是利用来自NIH SIG申请的资金以及相当大的机构支持实现的 来自MGH领导层。为我们的核心用户提供持续访问最新的共焦成像是一种 保持MGH社区处于NIH资助的与健康相关的研究的前沿的关键部分。