CVID Core Facilities
The Flow Cytometry Facility, directed by Dr. Mark Williams, provides investigators within the CVID the capability for multiparameter fluorescent analysis of any cultured or primary cells. Characteristics of sub-populations, based upon prototypical markers, in heterogeneous mixtures can easily be determined. Parameters typically analyzed in the facility include; expression of cell surface receptors/markers, levels of intracellular proteins including cytokines and fluorescent proteins (GFP), intracellular calcium flux or generation of reactive oxygen/nitrogen species, DNA content, cell proliferation, and multiple measures of cell death/apoptosis. The facility contains a BD FACSCalibur and a BD FACScan, with separate stations for analysis of data using CellQuest or FlowJo.
Contact: Mark Williams
The histology core is directed by Ms. Elizabeth Smith and provides services in necropsies and perfusion fixation of various research animal species (rodents, rabbit, etc), processing of tissues and cells, embedding for paraffin and frozen sections, microtomy and cryotomy sectioning, and both routine and special staining. Consultation service is also available to provide advice concerning tissue processing, embedding orientation, and routine or special staining. Optimization of antibodies against newly discovered antigens for immunohistochemistry on frozen and paraffin sections and on cultured cells is also available. Equipment in the facility includes a Renaissance automatic tissue processor, Leica EG 1160 embedding center, a microtome cryostat and rotary microtomes for paraffin sections. The core also houses a Nikon Eclipse E 400 microscope with DS camera.
The microscopy core includes several conventional light and fluorescent microscopes, a confocal fluorescence microscope (Nikon BioRad Radiance 2100), and a Zeiss Axiovert 200 MOT live cell imaging system with a temperature controlled perfusion chamber. The system also contains Stallion deconvolution software. Image analysis is provided by the Volocity 4-dimensional imaging software (Improvision). The confocal instrumentation is under the direction of Dr. Irina Mikhailenko.
Contact: Irina Mikhailenko
Protein Analysis Core
The protein analysis core provides services that include: protein purification, protein sequencing and mass spectrometry. The core is directed by Mr. Brian Hampton, and is equipped with an Applied Biosystems Procise 494 protein sequencer, a Thermo Finnigan LCQ Advantage ion trap mass spectrometer, and three HPLC systems capable of micro analytical to preparative scale separations of proteins and peptides. This facility gives CVID members access to leading edge technologies that allow the determination of protein primary structure and the identification of post-translational modifications.
Contact: Brian Hampton
Protein Interaction Core
The protein interaction core is equipped to assist CVID members in measuring and quantifying protein-protein interactions. This core includes an SLM fluorimeter, a recently acquired JASCO circular dichrometer, and a BiaCore 3000 surface plasmon resonance instrument. The core is under the direction of Dr. Leonid Medved. Recently, we have been successful in interfacing the BiaCore 3000 technology with a mass spectrometer to identify proteins captured on the BiaCore chip.
Contact: Leonid Medved