Research Core Facilities Rationale and Organization. We propose 3 core facilities (2 continuing and 1 new): 1) Cell Culture, 2) Microscopy and 3) Computation and Bioinformatics. Cell Culture and Microscope Cores are located in adjacent labs to provide seamless transition from cell culture to microscopy. All core facilities are for the exclusive use of Center members. CORE A: Cell Culture The essential role of this core is to standardize every aspect of the cell culture system to maximize efficiency, consistency, and accessibility. It will serve as a repository for all cell lines, vectors, and reagents (media, supplements, basement membrane extract). Dr. Lourdes Estrada will be the core PI, and daily operations will be performed by two staff with dedicated effort. CORE B: Microscopy This Core will run and maintain the adjacently located HCAM system and LSM 510 Scanning Confocal Microscope, provide general oversight of equipment, coordinate the scheduling of users, provide expertise and in depth training, and help ensure that Center members can efficiently generate necessary data for models models. Dr. Darren Tyson, an expert in advanced imaging technology, will be responsible for the operations. BD Pathway 855 Bioimager - The Bioimager is capable of imaging an entire 96-well microplate in a single channel and focal plane in ~8 min. Imaging can also be performed repeatedly with multiple images per well, multiple focal planes (z-sections), and using multiple fluorescent channels. In addition, automated liquid handling allows for drug treatments, with precise control of duration and volume. This machine has adaptable hardware that directly integrated with OME to expedite downstream processing of image data. Scanning Confocal Imaging System (LSM5 #28 405/AR+2HENE/3PMT 3.5) - This system is used for high-resolution image capturing particularly for 3D cultures. Its features include: the latest operating system and hardware, faster 3D rendering, a 405 nm """"""""UV-like"""""""" laser for DAPI and Hoechst, FRAP capability for photobleaching, photoactivation, and workstations for image processing. CORE C: Computational and Bioinformatics The Center needs for high-performance computing are three-fold: 1) implementation of computational models;2) acquisition of high-throughput quantitative cellular phenotypic parameters (at the single-cell level);and 3) image data usability via extensive annotation, processing and online databases.
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