HIGH RESOLUTION MICROSCOPY

高分辨率显微镜

• 批准号: 7414795
• 负责人: PETER D. EMANUEL
• 金额: $ 17.04万
• 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: 2007-05-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414795
• 关键词: Argon; Arts; Award; Biological; Biomedical Research; Cancer Center; Cell-Matrix Junction; Cells; Comprehensive Cancer Center; Computer software; Consultations; Data Analyses; Detection; Devices; Equipment; Fiber; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Foundations; Funding; Generations; Grant; Health Services; Helium; Image; Imaging Techniques; Imaging technology; Ions; Krypton; Laser Scanning Confocal Microscopy; Laser Scanning Microscopy; Lasers; Life; Light; Measurement; Medical; Medical center; Microscopy; National Center for Research Resources; Neon; Optics; Output; Phototoxicity; Preparation; Productivity; Range; Research; Research Personnel; Resolution; Resources; Scanning; Source; Specialist; Specimen; Structure; Surface; System; Techniques; Tissues; U-Series Cooperative Agreements; United States Department of Veterans Affairs; United States National Institutes of Health; Work; data acquisition; design; detector; digital; digital imaging; experience; fluorophore; improved; instrument; instrumentation; member; nanometer; research study; submicron; tool; vision science

DESCRIPTION (provided by applicant): 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. In 1998, Dr. Kent Keyser was the recipient of an NIH NCRR Shared Instrument Grant, a UAB Health Services Foundation award, and start-up funds from the UAB Office of the Provost, the Chairs of Departments in both the Medical and Academic Divisions of UAB, and Center Directors. These funds were used to establish a new facility, the High Resolution Imaging Facility. This imaging resource is equipped with a Leica Confocal Imaging Spectrophotometer TCS SP unit. It is 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 DIG 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, Cancer Center 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 Helium:Neon 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. The Facility offers Comprehensive Cancer Center members access to a comprehensive imaging facility with a full range of instrumentation and the expertise to use these resources productively.

描述（由申请人提供）：生物中分子的检测和成像 系统是生物医学研究中极其重要的工具。 传统的荧光显微镜是一种非常有用的技术，但受到焦平面外源发射造成的分辨率损失的限制。 共焦激光扫描显微镜通过用激光照射的针孔发出的光扫描样本并通过检测器针孔对发射的光进行成像来解决这个问题。 所有离焦荧光都会被针孔拒绝，从而在 X、Y 和 Z 轴上实现亚微米分辨率。 1998 年，Kent Keyser 博士获得了 NIH NCRR 共享仪器补助金、UAB 健康服务基金会奖以及 UAB 教务长办公室、UAB 医学和学术部门各系主任以及中心主任提供的启动资金。 这些资金用于建立一个新设施，即高分辨率成像设施。 该图像资源是 配备徕卡共焦成像分光光度计 TCS SP 装置。 它被配置为 使用相干 MIRA Ti:Sa 超快激光器进行多光子激发，输出波长可在 690-990 纳米之间选择。 这使得能够以最小的光毒性对活细胞和组织以及距组织表面 200-400 微米的结构进行成像。 SP 允许用户选择要传送到检测器的发射光谱的特定部分。 其结果是更高的荧光产量、减少串扰并提高图像质量。 该设施还包括配备光纤发射氪离子和氩离子激光器的奥林巴斯 Fluoview CLSM、传统荧光和 DIG 显微镜设备以及配备数字反卷积软件、输出和图像存储设备的工作站。 通过与伯明翰退伍军人管理局医疗中心达成的合作协议，癌症中心成员可以使用配备紫外激光器和紫外校正光学器件的徕卡 TCS SP 装置。 徕卡仪器配备三个或四个检测器通道、一个透射光通道以及氩离子、氪离子和氦：氖激光器，并且能够同时对多达三个荧光团进行成像。 根据与细胞粘附和基质研究中心以及视觉科学研究中心的合作协议，HRIF 现在提供最先进的 FRET 系统。 该设施为综合癌症中心会员提供 使用配备全套仪器的综合成像设施以及高效利用这些资源的专业知识。