COBRE: U NEL: CORE B: MICROSCOPY & BIOIMAGING

COBRE：U NEL：核心 B：显微镜

• 批准号: 8360527
• 负责人: Javier Seravalli
• 金额: $ 15.18万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360527
• 关键词: Biochemistry; Biology; Biophysics; Calorimetry; Cells; Centers of Research Excellence; Chemicals; Circular Dichroism; Collaborations; Coupled; Detection; Differential Scanning Calorimetry; Elements; Fluorescence; Fluorescence Spectroscopy; Funding; Grant; Human Resources; Imaging technology; In Vitro; Measurement; Metals; Methodology; Methods; Microscopy; Mission; Modeling; Monitor; National Center for Research Resources; Nebraska; Oxidation-Reduction; Plasma; Population; Principal Investigator; Process; Proteins; Reaction; Reader; Research; Research Infrastructure; Research Personnel; Research Support; Resources; Running; Sampling; Scanning; Services; Source; Spectrum Analysis; Students; System; Technology; Training; Transition Elements; United States National Institutes of Health; Universities; analytical ultracentrifugation; bioimaging; cost; detector; graduate student; in vivo; instrument; instrumentation; interest; macromolecule; mass spectrometer; rapid technique; small molecule

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The purpose of the Core is to provide service support for the research in this proposal, in particular the characterization of in vitro and in vivo processes by spectroscopy, biophysical methods, metal analysis, microscopy and imaging technologies. Much of the important information about redox reactions and redox-related processes can be monitored by these methodologies. Examples of spectroscopy and biophysical applications are EPR, circular dichroism (CD), fluorescence, differential scanning calorimetry (DSC) and isothermal calorimetry (ITC). Rapid methods are available for EPR, UV/visible and fluorescence spectroscopies. Elemental analysis provides information on the relations between element concentrations (transition metals and main elements) and redox processes within proteins and cells. The primary missions of the Microscopy and Bioimaging Core are: 1) To provide state-of-the-art microscopy and bioimaging instrumentation services to RBC researchers, non-RBC researchers, and outside investigators; 2) To help facilitate strong collaborations between RBC researchers and non-RBC researchers within and outside the University of Nebraska system; and 3) To provide high quality instrumentation training of all the pertinent methods to graduate students, postdoctoral students and other interested personnel. Spectroscopy/Biophysics Core The core is located in rooms E155 and N113B of the Beadle Center at the UNL Campus in Lincoln. In addition to Redox Biology Center users, the core has been used by investigators within the Biochemistry department, the UNL campus and academic users outside of UNL. The current instrumentation is composed of a Hi-Tech (Tkg Scientific, Ltd.) double mixing stopped flow equipped with single wavelength absorbance, diode array and fluorescence detectors plus a chemical quench apparatus; a Varian Eclipse (part of Agilent Technologies, Inc.) spectrofluorimeter with multi wavelength detection, single and multiple cuvet sample holders and a 96 well-plate sample reader. The instrument is capable of collecting excitation and emission spectra as well as it is suitable for fluorescence resonance transfer (FRETs) measurements; a circular dichroism double beam instrument from OLIS, Inc. Metal analysis capability comes from an Inductively Coupled Plasma/Mass Spectrometer (ICPMS, Agilent model 7500ce) equipped with collision/reaction cell plus an autosampler capable of handling up to 540 samples in 6 96-well plates per run. The biophysical instruments are an Isothermal calorimeter from Microcal for the study of macromolecule/macromolecule interactions and macromolecule/small molecule interactions, a differential scanning calorimeter (Microcal) for the study of macromolecule stability and an analytical Ultracentrifugation (Beckman) for the study of size distributions of macromolecule populations.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 核心的目的是为本提案中的研究提供服务支持，特别是通过光谱学、生物物理方法、金属分析、显微镜和成像技术表征体外和体内过程。关于氧化还原反应和氧化还原相关过程的许多重要信息可以通过这些方法进行监测。 光谱学和生物物理应用的例子有 EPR、圆二色性 (CD)、荧光、差示扫描量热法 (DSC) 和等温量热法 (ITC)。 快速方法可用于 EPR、紫外/可见光和荧光光谱。 元素分析提供有关元素浓度（过渡金属和主要元素）与蛋白质和细胞内氧化还原过程之间关系的信息。 显微镜和生物成像核心的主要任务是： 1) 为红细胞研究人员、非红细胞研究人员和外部研究人员提供最先进的显微镜和生物成像仪器服务； 2) 帮助促进内布拉斯加大学系统内外的 RBC 研究人员和非 RBC 研究人员之间的强有力合作；和 3) 向研究生、博士后和其他感兴趣的人员提供所有相关方法的高质量仪器仪表培训。 光谱学/生物物理学核心 核心位于林肯内布拉斯加州大学校园 Beadle 中心的 E155 和 N113B 室。 除了氧化还原生物学中心用户之外，该核心还被生物化学系、UNL 校园内的研究人员以及 UNL 以外的学术用户使用。 目前的仪器由配备单波长吸光度、二极管阵列和荧光检测器的高科技（Tkg Scientific, Ltd.）双混合停流以及化学淬灭装置组成； Varian Eclipse（安捷伦科技公司的一部分）分光荧光计，具有多波长检测、单个和多个比色皿样品架以及 96 孔板样品读取器。 该仪器能够收集激发和发射光谱，并且适用于荧光共振转移（FRET）测量； OLIS, Inc. 的圆二色性双光束仪器。金属分析能力来自于配备碰撞/反应池的电感耦合等离子体/质谱仪（ICPMS，安捷伦型号 7500ce）以及能够在 6 96- 时间内处理多达 540 个样品的自动进样器。每次运行的孔板。 生物物理仪器包括用于研究大分子/大分子相互作用和大分子/小分子相互作用的 Microcal 等温量热计、用于研究大分子稳定性的差示扫描量热计 (Microcal) 和用于研究尺寸分布的分析超速离心机 (Beckman)大分子群体。