Each of the projects of this PPG application utilize mouse models to assess innate immune cellfunction in infectious, inflammatory and autoimmune disease. As such, each project will rely onhistologic analysis of tissues from experimental animals to determine innate immune cell numbers,location, and function (such as cytokine secretion). Histologic analysis of lymphoid architectures inrelationship to the disease models being studied will also be a key method for each of the projects.Since the PPG envisions extensive sharing of animal resources, ideas and technologies, it is importantthan standardized methods of tissue preparation, immune cell staining and immunofluorescent detectionbe utilized by all four projects in the PPG. This will allow rational comparison of results between theprojects.The Histology Core will be run through Dr. Lowell's group (Project #3) and will perform thefollowing analyses: 1) standard tissue fixation, paraffin embedding, sectioning and histochemicalstaining; 2) frozen tissue embedding sectioning and immunohistochemical staining; 3) frozen tissueembedding with immunofluorescent staining and fluorescent cell localization; 4) Multiplex cytokineanalysis on serum, tissue and cell supernatant samples using Luminex technologies.Core personnel have extensive experience with all of the above methods, as described in thePreliminary data. The core is equipped with an OTF5000 cryostat for frozen tissue sectioning and astandard Leica microtomb for paraffin sections. Routine microscopy is done with an Eclipse TS100Fepi-fluorescent inverted microscope with Cool-Scan 3.0 mega pixel digital imagining system. Access tolarger upright Zeiss epi-fluorescent microscopes is available through the UCSF Immunology ImagingCore. This PPG core will also contain a staining station for routine histologic stains and will maintain abattery of mAbs for immunostaining. Multiplex bead based analysis of both cytokines, intracellularsignaling molecules and nucleic acids can be performed on the new BioPlex200 analyzer that will beavailable to PPG personnel.
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