High Resolution Microscopy Core

高分辨率显微镜核心

• 批准号: 7352464
• 负责人: KENT T KEYSER
• 金额: $ 7.69万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7352464
• 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。传统的荧光显微镜是一种非常有用的技术，但受到损失的限制。 由焦平面外的辐射源发射所引起的分辨率。共聚焦激光扫描 显微镜通过用来自激光照射的针孔的光扫描样品来避开这个问题。 并通过探测器针孔对发射的光成像。所有散焦的荧光都会被 针孔，导致在X、Y和Z轴上的亚微米分辨率。 C 1998年，来自NCRR共享仪器赠款、UAB卫生服务基金会奖和其他 资源来自UAB教务长办公室，医学院和学术界的科室主任 DAB的部门和中心主任被用来建立高分辨率成像设备。这 共享资源配备徕卡共焦成像分光光度计TCS SP单元，配置用于 输出可在690-990范围内选择的相干Mira Ti：Sa超快激光器多光子激发 纳米级这允许以最小的光毒性对活细胞和组织成像，以及对结构200- 距离组织表面400微米。SP允许用户选择排放的特定部分 光谱将被传送给探测器。其结果是更高的荧光产量，减少串扰，以及 改善了图像质量。该设施还包括一台奥林巴斯FluoView CLSM，带有光纤发射的氪 离子和Ar离子激光器、常规荧光和DIC显微镜设备以及工作站 配有数字反卷积软件、输出和图像存储设备。通过一个合作社 与伯明翰退伍军人管理医疗中心达成协议，RDCC成员可以访问 徕卡TCS SP单元配备了紫外光激光器和紫外光校正光学元件。徕卡仪器配备齐全 具有三个或四个探测器通道、一个透射光通道、以及Ar离子、氪离子和 氦-霓虹灯激光器，并且能够同时成像多达三个荧光团。在一个合作社里 与细胞黏附和基质研究中心以及视觉科学研究中心达成协议， HRIF现在提供对最先进的FRET系统的访问。为新的活体成像提供大量资金 通过成功的NIH Blueprint应用程序获得了工具，并共享了 关于高速图像采集能力的仪器授权申请正在审批中。HRIF服务了70年 RDCC调查员在本供资期间和他们的使用占总设施的49% 用法。在同一时期，RDCC P30机制提供的支持不到 该设施的年度运营总成本。