High Resolution Microscopy Core

高分辨率显微镜核心

基本信息

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
项目类别：
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

项目摘要

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之外的源发射引起的分辨率。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来解决这个问题并对通过探测器针孔发射的光进行成像。所有离焦荧光均被拒绝针孔，从而在 X、Y 和 Z 轴上实现亚微米分辨率。 c 1998 年，来自 NCRR 共享仪器补助金、UAB 健康服务基金会奖和其他来自 UAB 教务长办公室、医学和学术部门主席的资源 DAB 各部门和中心主任负责建立高分辨率成像设施。这共享资源配备了徕卡共焦成像分光光度计 TCS SP 单元，配置用于使用相干 MIRA Ti:Sa 超快激光器进行多光子激发，输出可在 690-990 之间选择纳米这允许以最小的光毒性对活细胞和组织以及结构进行成像 200- 距组织表面 400 微米。 SP 允许用户选择发射的特定部分光谱被传送到检测器。结果是更高的荧光产量、更少的串扰，以及提高图像质量。该设施还包括一台带有光纤发射氪的奥林巴斯 Fluoview CLSM 离子和氩离子激光器、传统荧光和 DIC 显微镜设备以及工作站配备数字反卷积软件、输出和图像存储设备。通过合作社与伯明翰退伍军人管理局医疗中心达成协议，RDCC 成员可以获得 Leica TCS SP 装置配备紫外激光器和紫外校正光学器件。徕卡仪器配备具有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 HeliunrNeon 激光器，能够同时对多达三个荧光团进行成像。在合作社与细胞粘附和基质研究中心、视觉科学研究中心、 HRIF 现在提供最先进的 FRET 系统。新的体内成像的大量资金仪器已通过成功的 NIH Blueprint 应用程序和共享高速图像采集功能的仪器资助申请正在等待中。 HRIF 服务了 70 当前资助期间RDCC研究人员及其使用量占设施总数的49% 用法。同期，RDCC P30机制的支持占比不足20% 设施的年度运营总成本。