The Microscopy and Flow Cytometry Core (MFCC) of the COBRE Center for Targeted Therapeutics (CTT) provides services to all four CTT research projects and other investigators supported by the CTT and other IDeA programs at the University of South Carolina (USC). These services include training in microscopy and flow cytometry, advice on study design, with an emphasis on the analysis of cellular drug effects, and assistance with the instrumentation usage, both at the Core and at other USC facilities (in particular at the USC School of Medicine), as well as expert assistance in anatomic pathology of human and animal tissues. The available equipment includes: Carl Zeiss LSM700 Laser scanning confocal microscope with 4 lasers functioning from 405 to 633 nm, equipped for live cell imaging (with environmental chamber), standard point scanning, 3D reconstruction, FRAP, FRET, time lapse and tile/stitching; a Leica Microsystems AS MDW fluorescence microscope optimized for live cell imaging with temperature and CO2 control; two Leica Microsystems DM IRE2 fluorescent microscopes, which are motorized for z-stack acquisition and one of which has an environmental chamber. Olympus IX81 fluorescence microscope with workstation running Metamorph software is used for fluorescence imaging and deconvolution of wide field fluorescence images; it also has a Sutter Instruments Smartshutter and rapid filter changing wheels for ratiometric measurement of intracellular ion changes. There are 3 Carl Zeiss Axiovert 200 microscopes and an Olympus upright BX41 microscope, each with a color camera. All microscopes are in dedicated rooms and on vibration isolation tables. Flow cytometric equipment includes the BD FACS ARIA III for cell sorting, BD LSR II Flow Cytometer with high-throughput option and a 96-well plate loader, and Beckman Coulter FC500 flow cytometer. Users may operate the microscopes and one of the flow cytometers independently, after training by the co-directors. Many standard protocols have been established for immunohistochemistry and immunofluorescence analysis of cells and tissues, and for the analysis of cell growth, cell death and cell cycle progression. The core also provides services of an experienced anatomic pathologist, with a broad experience in gross pathology and histopathology in human and animal studies. The pathology core services include advice on study design and specimen processing including paraffin and cryosectioning, routine H&E staining, histochemical stains, and immunohistochemical analysis. Pathology reports will include macroscopic and microscopic findings, diagnosis of lesions, discussion of significant findings, and biological significance of the pathological effects of genetic modifications and experimental compounds in different animal organs, as well as tumor morphology in animal models and in human tissue samples. The core personnel offer advice on the choice of instruments and experimental protocols, provide training and assist with preparation of manuscripts and grant proposals.
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