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.

项目概要 我们请求资金来支持我们现有的 Nikon A1R 共焦显微镜的升级，该显微镜现已 已有近 10 年历史，缺乏许多自购买以来开发的新功能 2012. 升级后的仪器将安装在膜生物学 (PMB) 显微镜计划内 麻省总医院 (MGH) 多学科 Simches 研究中心的核心 建筑。这个经 MGH 批准的核心设施的任务是提供全方位的显微镜检查 向 2 个 NIH 资助的 P30 中心和 MGH 社区内的各个研究小组提供技术，以及 超过。该核心在我们使用的静态和活细胞成像技术方面拥有丰富的专业知识 多年来拥有广泛的用户基础。四年前的收购增强了我们的成像能力 Zeiss LSM800 Airyscan 共焦平台可与旧款 Nikon A1R 一起使用。一起这些 仪器已提供足够的高水平成像带宽来服务我们的用户。尼康 A1R 是 广泛用于长期活细胞成像，并具有倒置显微镜和环境 室来满足这一需求。 LSM800，与正置显微镜关联，无需环境 室，提供一致且及时的访问高分辨率共焦，主要用于执行 固定样本的静态成像。随着表达各种活细胞成像的需求增加 荧光分子和标记蛋白质的类型，以及随着时间的推移对这些细胞成像的需求不断增加 使用 3D 重建，该应用程序的目的是为我们的用户提供现代的访问权限 共焦显微镜选项比当前较旧的选项更适合专门的成像目的 A1R系统。特别是，新的高速高清共振扫描模式与相关的AI图像 与传统的处理能力相比，图像尺寸和分辨率显着增加 旧的共振扫描系统。再加上用新的 GaAsp 替换旧的 PMT 探测器 探测器和稳定固态激光器的结合，这些升级将使样本能够 暴露于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，分辨率显着提高。此外，图像和多色 3D 堆栈 在共振扫描模式下可以更快地收集，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 多年来已支持数百份同行评审出版物和数十项资助申请。这 Core 不断寻求不断改进和升级 Core 设施以支持其用户群。 这是通过 NIH SIG 申请的资金以及大量机构支持实现的 来自 MGH 领导层。为我们的核心用户提供持续访问最新共焦成像的能力是 这是维持 MGH 社区在 NIH 资助的健康相关研究前沿的重要组成部分。