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.
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