Olympus TIRF Microscope for Simultaneous Multichannel Live cell Imaging

用于同时多通道活细胞成像的奥林巴斯 TIRF 显微镜

• 批准号: 8447267
• 负责人: RADU VIRGIL STAN
• 金额: $ 17.96万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2014-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447267
• 关键词: Aging; Americas; Biochemical; Biochemistry; Biological; Biological Sciences; Cell membrane; Cell physiology; Cells; Communities; Complement; Comprehensive Cancer Center; Computer software; Core Facility; Coupled; Cytoskeletal Modeling; Endocytosis; Ensure; Fluorescence; Funding; Goals; Gossypium; Immunology; Laboratories; Lasers; Life; Maintenance; Manuals; Medicine; Membrane Protein Traffic; Microbiology; Microscope; Microscopy; Molecular; Optics; Pathology; Phototoxicity; Recovery; Research Personnel; Resolution; Schools; Speed; System; Technology; United States National Institutes of Health; Work; cellular imaging; college; instrument; medical schools; optical imaging; particle; pathogen; photoactivation

DESCRIPTION (provided by applicant): A diverse group of 6 NIH funded investigators and a junior investigator within Departments of Pathology, Biochemistry, Medicine, Microbiology and Immunology at Dartmouth Medical School and Biological Sciences at Dartmouth College request funds to purchase a state of the art multicolor TIRFM live cell imaging system from Olympus America. The system integrates an Olympus IX-81 inverted microscope, an automated multicolor total internal reflection microscopy (TIRFM) illuminator (Olympus Celltirf) equipped with three lasers 405/491/561, and capable of FRAP/ photoactivation; an ultra-fast/highly sensitive sCMOS camera (Andor Neo) coupled to a DualView beam splitter; fully integrated MetaMorph for Olympus acquisition/analysis software optimized for 1) rapid/ultra-sensitive 4D four channel acquisition; 2) FRAP and photoactivation and 3) particle tracking; The new state-of-the-art workstation will complement our heavily utilized Zeiss LSM 510 META and aging single channel manual TIRFM system, while constituting a significant upgrade in multi channel acquisition, speed and sensitivity. Importantly, the multicolor TIRFM system will provide new/unique total internal fluorescence capabilities for the Optical Cellular Imaging Facility that s utilized regularly by multiple schools and departments comprising >75 laboratories and ~300 users. Our investigators, all well funded through the NIH, work on a host of cell biological and biochemical questions aimed at understanding molecular mechanisms of fundamental cellular processes such as membrane trafficking, endocytosis, pathogen infectivity and cytoskeletal organization and interaction with the plasma membrane. Satisfactory completion of the stated goals of the NIH-sponsored projects of these users requires examination of dynamic cellular processes in live cells with high spatial and temporal resolution and reduced phototoxicity. The total internal reflection microscope described in this proposal is a state of the art instrument tht satisfies these needs by using proven technology. In the requested configuration, the multicolor TIRFM workstation will afford our disparate group of investigators a broad range of basic/advanced live-cell microscopy capabilities that are presently deficient/absent at Dartmouth. It would represent the only instrument of its kind to serve the entire Dartmouth life science community. The new system will be conveniently located, administered, and maintained, in the Norris Cotton Comprehensive Cancer Center Optical Imaging Core Facility (OICF); and it will be incorporated into the OICF's existing, well established recharge system to ensure recovery of funds for supplies and maintenance.

描述（由申请人提供）：达特茅斯医学院的病理学，生物化学，医学，微生物学和免疫学部门的一组由6名NIH资助的研究人员和一名初级调查员，达特茅斯学院的生物学科学，要求从Olympus向Olympus购买ART Multicolor Tirfm Live Tirfm Live Tirfm Live Mive System。该系统集成了Olympus IX-81倒置显微镜，这是一种自动多色总内反射显微镜（TIRFM）照明器（Olympus CellTirf），配备了三个激光器405/491/561，并且能够使用FRAP/Photoactivation；超快速/高度敏感的SCMOS摄像头（Andor Neo）与双视梁分离器相连；针对奥林巴斯采集/分析软件进行了完全集成的Metamorph，针对1）快速/超敏感的4D四通道采集； 2）FRAP和光激活以及3）粒子跟踪；新的最先进的工作站将补充我们大量利用的Zeiss LSM 510 meta和衰老的单通道手动TIRFM系统，同时构成了多通道获取，速度和灵敏度的重大升级。重要的是，多色TIRFM系统将为光学蜂窝成像设施提供新/独特的总内荧光功能，该功能由多个学校和部门定期使用，包括> 75个实验室和约300位用户。我们的研究人员都通过NIH提供了充分的资金，致力于许多细胞生物学和生化问题，旨在理解基本细胞过程的分子机制，例如膜运输，内吞作用，病原体感染性和细胞骨架组织以及与质膜的相互作用。满足这些用户NIH赞助项目的既定目标的令人满意的完成需要检查具有高空间和时间分辨率和降低光毒性的活细胞中的动态细胞过程。本提案中描述的总内反射显微镜是通过使用验证的技术来满足这些需求的最先进的状态。在请求的配置中，多色TIRFM工作站将为我们不同的研究人员提供多种基本/高级活细胞显微镜功能，这些功能目前在达特茅斯缺乏/缺乏。它将代表为整个达特茅斯生活科学界服务的唯一同类工具。新系统将在诺里斯棉花综合癌症中心光学成像核心设施（OICF）中方便地，管理和维护；它将被整合到OICF现有的，良好的充值系统中，以确保收回用于供应和维护的资金。