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Core C: Kidney Imaging
The Kidney Imaging Core is a national resource that provides qualitative and quantitative image analysis of the kidney and lower urinary tract. These studies are performed using microscopic techniques specifically developed to image these tissues, and support other Center Cores that use physiological and molecular approaches to understand the function and dysfunction of the kidney and lower urinary tract. The objectives of the core are:

• To perform light and electron microscopic analysis of epithelial cells and tissues. These services are performed by the Epithelial Imaging Subcore, which specializes in the imaging and analysis of cultured kidney- and bladder-derived epithelial cells, as well as kidney and lower urinary tract-associated tissues and tissue slices. Imaging capture modalities include brightfield, DIC, darkfield, confocal (employing either single pinhole or spinning disk technologies), as well as STED microscopy. For samples that show weak fluorescence, a video camera system is available with associated digital deconvolution software. Analysis includes colocalization and tracking in 4D, as well as 3D reconstruction of individual epithelial cells and tissues using Imaris, Metamorph, and Volocity imaging softwares. Furthermore, the University of Pittsburgh Center for Biological Imaging (CBI), will provide specialized imaging tools and analysis including two-photon laser scanning microscopy of organs/tissues, fluorescence-resonance energy transfer (FRET), fluorescence recovery after photobleaching (FRAP), Ca2+ imaging, and high-speed total internal reflection fluorescence microscopy (TIR-FM). Ultrastructural analyses include scanning electron microscopy (SEM), and transmission electron microscopy (TEM), including ultrathin immunoelectron microscopy to examine the distribution and relationship of single or multiple antigens.

• To quantify and reconstruct kidney and lower urinary tract-associated tissues and organs in 3D. The Stereology and 3D Organ Reconstruction Subcore will provide image analysis that includes, but is not limited to stereology, a set of unbiased and accurate tools to measure parameters such as volume, surface area, length, and number. In addition, this subcore, along with the Epithelial Imaging Subcore, will provide 3D reconstructions of cells, tissues, and organs using Imaris, Neuroclida, Metamorph, Volocity, and StereoImager software packages.

• To provide expert analysis of tissue and organ histology and pathology. The Histology/Pathology Subcore specializes in the preparation of kidney and lower urinary tract tissues for histology and electron microscopy, and subsequent analysis of pathological findings by our expert kidney/lower pathologist.

• To visualize and quantify molecular and cellular dynamics in living animals. The Live-Animal Imaging Subcore is dedicated to intravital microscopy of kidneys in normal and pathological states. Imaging is provided by a dedicated two-photon microscope system as well as by the CBI, which has additional two-photon setups.

• To provide training in the processing, staining, and imaging of kidney and lower urinary tract cells and tissues as well as interpretation of morphological data. To enhance the likelihood of successful morphological studies, the Core will provide advice and instruction in the design of these experiments. Beyond providing technical expertise, a crucial function of the Imaging Core will be to train investigators in the proper fixation, processing, and imaging of kidney and lower urinary tract tissues as well as training on the specialized instruments available to core users.