COBRE: U NEL: CORE B: MICROSCOPY & BIOIMAGING

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 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。许多关于氧化还原反应和与氧化还原相关的过程的重要信息都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电子顺磁共振、圆二色谱(CD)、荧光、差示扫描量热法(DSC)和等温量热法(ITC)。电子顺磁共振、紫外可见光谱和荧光光谱都有快速方法可用。元素分析提供了元素浓度(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的关系的信息。 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州大学系统内外的红细胞研究人员和非红细胞研究人员之间的密切合作；以及 3)为研究生、博士后和其他感兴趣的人员提供各种相关方法的高质量仪器培训。 光谱学/生物物理学核心 核心位于林肯UNL校区执事中心的E155和N113B房间。除了氧化还原生物中心的用户外，核心还被生物化学系、UNL校园内的调查人员和UNL以外的学术用户使用。目前的仪器是由一个高科技(TKG Science，Ltd.)配备了单波长吸收、二极管阵列和荧光检测器以及化学猝灭装置的双混合停止流；Varian Eclipse(安捷伦技术公司的一部分)具有多波长检测功能的荧光分光光度计、单个和多个CUVE样品架和96孔板样品读取器。该仪器能够收集激发光谱和发射光谱，并适用于荧光共振转移(FRET)测量；OLIS，Inc.的圆二色双光束仪器。金属分析能力来自配备碰撞/反应池的电感耦合等离子体/质谱仪(ICPMS，Agilent型号7500ce)以及能够在每次运行中处理6 96孔板中540个样品的自动进样器。这些生物物理仪器是MicroCal的等温量热仪，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，差示扫描量热仪(MicroCal)用于研究大分子的稳定性，分析超速离心仪(Beckman)用于研究大分子群体的尺寸分布。

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.