The Cell Morphology Core (CMC) supports the grant members with all aspects of histology-related work necessary to accomplish their research goals. The core is directed by Dr. Peter Bell and is housed in 800 square feet of dedicated space. It owns all the equipment necessary to process tissues and cells for a wide array of histological analyses. This includes several fluorescence microscopes, a transmission electron microscope, microtomes, an ultramicrotome, several cryostats, and automated tissue processing equipment. For tissue analysis at the light microscope level, paraffin, frozen and plastic-embedded samples are used. Routine and special stains as well as the detection of reporter gene expression in tissues (GFP, LacZ) are performed regularly. Further techniques include immunostaining, in situ hybridization, apoptosis and cell proliferation assays. Immunostaining is performed with fluorescent or enzyme-labeled antibodies, using all the appropriate positive and negative controls. In situ hybridization for identification of transduced cells or the detection of viruses uses labeled DMA or RNA probes. Images can be acquired by widefield or confocal microscopy and analyzed with Image-Pro Plus and AnalySIS software, quantifying the degree of staining for any target. Transmission electron microscopy (TEM) is performed to evaluate tissue samples at an ultrastructural level. For vector analysis, negative staining of viral preparations is applied for EM analysis. Viral vectors are assessed for average density, purity, uniformity and viral particle integrity. Contaminations in adenovirus preparations with AAV (and vice versa) are detectable due to the difference in size of both viruses. Investigators can also receive training in histological techniques and microscopy allowing them to perform their own analyses by utilizing the core's resources.
The Cell Morphology core processes and analyzes tissues both for histopathology and the expression of transgenes, important parameters to assess the success of gene therapy studies. The histological examination of tissues is aimed to detect how gene transfer experiments affect target tissues and other organs.
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