Cores

研究核心设施 基本原理和组织结构。我们提出了3个核心设施(2个仍在使用，1个新的)：1)细胞培养，2)显微镜，3)计算和生物信息学。细胞培养和显微镜核心位于相邻的实验室，以提供从细胞培养到显微镜的无缝过渡。所有核心设施均为中心会员专用。 核心A：细胞培养 这个核心的基本作用是标准化细胞培养系统的每个方面，以最大限度地提高效率、一致性和可及性。它将作为所有细胞系、载体和试剂(介质、补充剂、基膜提取物)的储存库。Lourdes Estrada博士将是核心PI，日常手术将由两名工作人员专心致志地执行。 核心B：显微镜 该核心将运行和维护相邻的HCAM系统和LSM 510扫描共焦显微镜，提供设备的一般监督，协调用户的调度，提供专业知识和深入培训，并帮助确保中心成员能够有效地为模型生成必要的数据。先进成像技术专家达伦·泰森博士将负责手术。BD Path 855生物成像仪-该生物成像仪能够在~8分钟内在单通道和焦平面内对整个96孔微孔板进行成像。还可以利用每个井的多个图像重复执行成像， 多个焦平面(z截面)，并使用多个荧光通道。此外，自动化液体处理允许药物治疗，并精确控制持续时间和体积。该机器具有可适应的硬件，可直接与OME集成，以加快图像数据的下游处理。扫描共焦成像系统(LSM5#28 405/AR+2HENE/3PMT 3.5)-该系统用于高分辨率图像捕获，特别是3D培养。它的功能包括：最新的操作系统和 硬件、更快的3D渲染、用于DAPI和Hoechst的405 nm“类UV”激光器、用于光漂白、光激活的FRAP功能以及用于图像处理的工作站。 核心C：计算和生物信息学 该中心对高性能计算的需求有三个方面：1)计算模型的实施；2)高通量定量细胞表型参数的获取(在单细胞水平上)；以及3)通过广泛的注释、处理和在线数据库获得图像数据的可用性。

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.