High Resolution Microscopy Core

高分辨率显微镜核心

• 批准号: 7669291
• 负责人: KENT T KEYSER
• 金额: $ 9.49万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7669291
• 关键词: Accounting; Applications Grants; Argon; Arts; Award; Biological; Biological Markers; Biomedical Research; Cell-Matrix Junction; Cells; Center Core Grants; Computer software; Consultations; Data Analyses; Detection; Development; Devices; Equipment; Evaluation; Extramural Activities; Fiber; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Foundations; Functional disorder; Funding; Generations; Grant; Health Services; Image; Imaging Techniques; Imaging technology; In Vitro; Ions; Krypton; Laser Scanning Confocal Microscopy; Laser Scanning Microscopy; Lasers; Life; Light; Measurement; Medical; Medical center; Microscopy; Mission; National Center for Research Resources; Neurosciences; Optics; Other Resources; Output; Pattern; Phototoxicity; Preparation; Productivity; Reagent; Research; Research Personnel; Resolution; Resource Sharing; Resources; Rheumatism; Role; Scanning; Source; Specialist; Specificity; Specimen; Speed; Staging; Structure; Support of Research; Surface; System; Technical Expertise; Techniques; Testing; Tissues; U-Series Cooperative Agreements; United States Department of Veterans Affairs; United States National Institutes of Health; Work; cost; data acquisition; design; detector; digital; digital imaging; experience; fluorophore; improved; in vivo; instrument; instrumentation; member; mouse model; nanometer; novel; research study; submicron; tool; vision science

The detection and imaging of molecules in biological systems is a vitally important tool in biomedical research. Conventional fluorescence microscopy is an extremely useful technique, but is limited by the loss of resolution caused by emission from sources outside of the focal plane. Confocal Laser Scanning Microscopy circumvents this problem by scanning the specimen with light from a laser-illuminated pinhole and imaging the emitted light through a detector pinhole. All out-of-focus fluorescence is rejected by the pinhole, resulting in submicron resolution in the X, Y, and Z axes. c In 1998, funds from a NCRR Shared Instrument Grant, a UAB Health Services Foundation award, and other resources from the UAB Office of the Provost, the Chairs of Departments in both the Medical and Academic Divisions of DAB, and Center Directors were used to establish, the High Resolution Imaging Facility. This shared resource is equipped with a Leica Confocal Imaging Spectrophotometer TCS SP unit configured for multiphoton excitation with a Coherent MIRA Ti:Sa ultrafast laser with output selectable between 690-990 nanometers This permits imaging of living cells and tissues with minimal phototoxicity, and of structures 200- 400 micrometers from the tissue surface. The SP allows user selection of specific portions of the emission spectrum to be delivered to the detector. The result is higher fluorescence yield, reduced crosstalk, and improved image quality. The Facility also includes an Olympus Fluoview CLSM with fiber launched Krypton ion and Argon ion lasers, conventional fluorescence and DIC microscopy equipment, and work stations equipped with digital deconvolution software, output and image storage devices. Through a cooperative agreement with the Birmingham Veteran's Administration Medical Center, RDCC members have access to a Leica TCS SP unit equipped with a UV laser and UV corrected optics. The Leica instruments are equipped with either three or four detector channels, a transmitted light channel, and Argon ion, Krypton ion and HeliunrNeon lasers, and are able to image up to three fluorophores simultaneously. In a cooperative agreement with the Cell Adhesion and Matrix research center, and the Vision Science Research Center, the HRIF now offers access to a state-of-the-art FRET system. Significant funds for new in vivo imaging instrumentation have been obtained through a successful NIH Blueprint application, and a Shared Instrument Grant application for high speed image acquisition capability is pending. The HRIF served 70 RDCC investigators during the current funding period and their use accounted for 49% of the total facility usage. During this same period, support from the RDCC P30 mechanism accounted for less than 20% of the total annual operating costs of the facility.

生物系统中分子的检测和成像是生物医学中至关重要的工具 研究。常规的荧光显微镜是一种非常有用的技术，但受损失的限制 由焦平面以外的来源发射引起的分辨率。共聚焦激光扫描 显微镜通过用激光释放的针孔扫描样品来避免此问题 并通过检测器针孔成像发出的光。所有聚焦荧光都被 针孔，导致X，Y和Z轴中的亚微分辨率。 c 1998年，来自NCRR共享工具赠款，UAB Health Services Foundation奖和其他的资金 来自教务长的UAB办公室的资源，医学和学术的部门主席 使用DAB和中心董事的划分是建立高分辨率成像设施。这 共享资源配备了Leica共聚焦成像分光光度计TCS SP单元配置为 具有连贯的mira ti：SA超快激光器的多光子激发，可在690-990之间选择输出 纳米允许对活细胞和组织进行最小的光毒性和200--结构的成像。 来自组织表面的400微米。 SP允许用户选择排放的特定部分 要传递到检测器的光谱。结果是更高的荧光产量，串扰降低，并且 改进的图像质量。该设施还包括带有纤维的Olympus Fluoview CLSM 离子和氩离子激光器，常规荧光和DIC显微镜设备以及工作站 配备了数字反卷积软件，输出和图像存储设备。通过合作 与伯明翰退伍军人行政医疗中心协议，RDCC成员可以访问 配备有紫外线和紫外线校正光学的Leica TCS SP单元。 Leica仪器配备了 具有三个或四个探测器通道，一个透射光通道和氩离子，k k离子和 Heliunrneon激光器，能够同时最多图像三个荧光团。在合作社 与细胞粘附和矩阵研究中心以及视觉科学研究中心的一致 HRIF现在提供了对最先进的品格系统的访问。新的体内成像的大量资金 通过成功的NIH蓝图应用程序获得了仪器，并共享 仪器赠款申请高速图像采集能力正在等待。 HRIF服务70 RDCC调查人员在当前的资金期间，其使用占总设施的49％ 用法。在同一时期，RDCC P30机制的支持占不到20％ 该设施的年度运营成本。