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; 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，现在 自购买以来，将近10岁，缺少许多新功能 2012年。升级的仪器将安装在膜生物学（PMB）显微镜的程序中 马萨诸塞州综合医院（MGH）的核心 建筑。该MGH批准的核心设施的任务是提供全范围的显微镜 MGH社区内的2个NIH资助的P30中心和个人研究小组的技术以及 超过。核心在我们使用的静态和活细胞成像技术方面具有相当大的专业知识 多年来，广泛的用户基础。 4年前，我们的成像毛细管增强了 Zeiss LSM800 Airyscan共聚焦平台与较老的Nikon A1R一起运行。在一起 仪器提供了足够的高级成像带宽，可以为我们的用户提供服务。尼康A1R是 广泛用于长期，活细胞成像，具有倒置显微镜和环境 会议室以促进这一需求。 LSM800，与直立显微镜相关，没有环境 会议厅，提供一致且及时的访问高分辨率共介焦，主要用于执行 固定标本的静态成像。随着表达各种活细胞成像的成像需求的增加 荧光分子和标记的蛋白质的类型，随着时间的推移，越来越多的需要对这些细胞进行成像 使用3D重建，此应用程序的目的是为我们的用户提供对现代的访问 共聚焦显微镜选项比当前较老更好地适合专业成像目的适应 A1R系统。特别是，具有相关AI图像的新型高速高清共振扫描模式 处理功能可在兼容到该功能时的图像尺寸和分辨率显着增加 旧的谐振扫描系统。再加上旧PMT探测器的新GAASP再替换 探测器和稳定固态激光器的掺入，这些升级将允许样品为 暴露于较低的激光强度，减少光漂白和光损伤，同时提供 与旧系统相比，分辨率显着增加。此外，图像和多色3D堆栈 可以在谐振扫描模式下更快地收集，同时又不牺牲质量，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，已经支持数百份经同行评审的出版物和数十种赠款申请。这 Core持续不断地改善和升级核心设施以支持其用户群。 这是使用NIH SIG应用程序的资金以及大量机构支持来实现的 来自MGH领导。向我们的核心用户提供持续访问最新的共聚焦成像是一个 维持MGH社区处于NIH资助，与健康相关的研究的最前沿的重要组成部分。