MRCE Live Cell Microscopy Core

MRCE 活细胞显微镜核心

基本信息

批准号：
7672138
负责人：
Haibing Teng
金额：
$ 18.44万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-09 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7672138
关键词：
Biohazardous Substance Biological Cells Communities Confocal Microscopy DNA Disease Disease Outbreaks Emergency Situation Emerging Communicable Diseases Environment Equipment Event Fluorescence Microscopy Fluorescence Resonance Energy Transfer Funding Gases Humidity Image Incubators Individual Infectious Diseases Research Investments Ions Life Light Maintenance Microbe Microscope Microscopy Operative Surgical Procedures Organelles Organism Photobleaching Photophobia Phototoxicity Preparation Proteins Protocols documentation Public Health RNA Recruitment Activity Research Research Personnel Safety Sampling Scientist Speed Techniques Technology Temperature Time Training Universities Washington Work biodefense biosafety level 3 facility combat cooking cost effective data acquisition design experience fluorescence microscope instrumentation laser capture microdissection member pandemic disease pathogen public health emergency response trafficking

项目摘要

This proposal describes the Live Cell Microscopy Core for the Midwest Regional Center of Excellence for Biodefense and Emerging Infectious Disease Research (MRCE). This Core is to provide MRCE, other regional centers and greater research community the ability to perform a variety of advanced fluorescence microscopy techniques with live BSL2 and BSL3 organisms. Before the facility was available, there was no regional option for performing advanced microscopy techniques on BSL3 organisms. The technical instrumentation and scientific expertise are also readily available to the public health community in the event of a bioterror, outbreak or pandemic event. Two fluorescence microscopes and a Laser-Capture Microdissection (LCM) microscope are available in the facility. The Nikon LiveScan Swept-Field Confocal microscope supports multi-dimensional data acquisition (x, y, z, time, wavelengths and multi-point), and the TILL Wide-Field microscope allows studies of ratio imaging (ions and pH) and FRET (Fluorescence Resonance Energy Transfer). Their super light sensitivity and video speed minimize photobleaching and phototoxicity, which is crucial for long-term imaging of fluorescent live samples. Cells are kept viable in a microscope cage-incubator that enables control of temperature, humidity and gas concentrations. Such livemotion imaging can be applied to explore dynamic localization, colocalization, trafficking, interaction and turnover between pathogen and cells, cellular organelles, proteins or ions. The LCM microscope enables detailed examination of extracted DMA, RNA and protein from targeted portions of histological sections. Perspective users will apply to the Core, and specific experimental protocols will be designed by the user in conjunction with the core PI. Biohazard safety issues will be resolved in conjunction with Dr. Susan Cook of Environmental Safety at Washington University, and appropriate clearances will be obtained in advance. Continued funding of this Core is required to increase the availability of cutting-edge microscopy for investigators in research community and in the event of an emergency to greatly advance our understanding biological mechanisms of pathogens.

该提案描述了中西部区域卓越中心的活细胞显微镜核心生物防御和新发传染病研究（MRCE）。这个核心是提供MRCE，其他区域中心和更大的研究团体执行各种先进荧光的能力活 BSL2 和 BSL3 生物体的显微镜技术。在该设施投入使用之前，没有对 BSL3 生物体执行先进显微镜技术的区域选项。技术方面在事件发生时，公共卫生界也可以随时获得仪器和科学专业知识生物恐怖、爆发或大流行事件。两台荧光显微镜和一台激光捕获仪设施内提供显微切割 (LCM) 显微镜。尼康 LiveScan 扫描场共焦显微镜支持多维数据采集（x、y、z、时间、波长和多点）， TILL 宽视野显微镜可用于比率成像（离子和 pH）和 FRET（荧光共振能量转移）。其超强的感光度和视频速度可最大限度地减少光漂白和光毒性，这对于荧光活体样品的长期成像至关重要。细胞保持活力显微镜笼式培养箱，可控制温度、湿度和气体浓度。这样的动态成像可用于探索动态定位、共定位、运输、交互和病原体与细胞、细胞器、蛋白质或离子之间的转换。 LCM 显微镜使对从组织切片目标部分提取的 DMA、RNA 和蛋白质进行详细检查。 Perspective用户将申请到Core，具体的实验方案将由用户设计与核心 PI 结合。生物危害安全问题将与苏珊·库克博士一起解决华盛顿大学的环境安全，并将提前获得适当的许可。需要继续资助该核心，以增加尖端显微镜的可用性研究界的调查人员以及在紧急情况下极大地增进我们的理解病原体的生物学机制。