Biophysical Imaging

摘要： 生物物理成像核心的主要目标是为PPG调查人员提供完整的范围 专业知识、培训、设备和数据分析工具，以获得纳米到微米尺度的生物物理信息 关于内皮细胞(EC)细胞骨架重排和屏障的细胞和分子基础的 监管。核心研发人员和设备将允许PPG科学家对3D结构进行成像，评估物理- CAL和化学性质，并定义微扰诱导的结构和活性的实时变化 细胞和亚细胞成分，包括膜、细胞骨架网络、细胞-基质和细胞-细胞连接-- 特兹。这个生物物理成像核心支持PPG的四个定量显微镜研究项目 与原子力显微镜(AFM)、共聚焦和去卷积荧光显微镜有关，全内 反射荧光(TIRF)显微镜、动态(实时)荧光成像、机械测量 细胞(牵引力显微镜)、超分辨率显微镜和自动化高通量定量检测。 显微镜可以观察细胞器(例如，皮质细胞骨架、连接复合体、焦点广告)。 肝和板脂)非常详细(3D)。核心D还将与每个项目和核心B(GE- 基因组/遗传学和蛋白质组核心)来培养经工程处理以表达不同nmMLCK1/2的内皮细胞， 皮质素，NAMPT，TLR4，DOCK1，层脂素，整合素β4，Kindlin-2，PSGL1，P-选择素和S1PR1/3受体 具有特定SNP或翻译后修饰突变的TORS转基因并测量各种生物物理 部队(TFM和AFM)。核心还提供对UNI的完整资源的经验使用- 包括最先进的高分辨率荧光显微镜在内的亚利桑那州成像核心设施的Verthy 系统。该核心还将利用亚利桑那州研究实验室的公共资源。 由Carol C.Gregorio博士领导的生物物理成像核心包括用于 大范围动态荧光成像、制造和机械生物物理学。核心D人员具有亲和力- 在高分辨率成像和物理化学检测领域的专业经验 性能，包括机械生物物理学和各种扫描探针显微镜。所有核心D实验 将在图森的亚利桑那大学进行。

ABSTRACT: The principal objective of the Biophysical Imaging Core is to provide PPG investigators with a complete range of expertise, training, equipment, and data analysis tools to obtain nano-to-micro scale biophysical information pertaining to the cellular and molecular basis of endothelial cell (EC) cytoskeletal rearrangements and barrier regulation. Core D personnel and equipment will allow PPG scientists to image 3D structures, evaluate physi- cal and chemical properties and define perturbant-induced real-time changes in the structures and activity of cells and subcellular constituents, including membranes, cytoskeletal networks, cell-matrix and cell-cell junc- tions. This Biophysical Imaging Core supports the PPG's four research projects with quantitative microscopy related to atomic force microscopy (AFM), confocal and deconvolution fluorescence microscopy, total internal reflection fluorescence (TIRF) microscopy, dynamic (live) fluorescence imaging, mechanical measurements of cells (traction force microscopy), super-resolution microscopy and automated high-throughput quantitative mi- croscopy to allow observation of cellular organelles (e.g., cortical cytoskeleton, junctional complexes, focal ad- hesions and lamellipodia) in great detail (3D). Core D will also interact with each Project and with Core B (Ge- nomic/Genetic and Proteome Core) to culture endothelial cells engineered to express different nmMLCK1/2, cortactin, NAMPT, TLR4, DOCK1, lamellipodin, integrin β4, kindlin-2, PSGL1, P-Selectin and S1PR1/3 recep- tors transgenes with specific SNP or post-translational modification mutations and measure various biophysical forces (TFM and AFM). The Core also offers access to experienced use of the complete resources of the Uni- versity of Arizona Imaging Core Facilities that includes state-of-the-art high-resolution fluorescence microscopy systems. This Core will also make use of the common resources available at Arizona Research Laboratories. The Biophysical Imaging Core, led by Carol C. Gregorio, PhD, includes personnel and laboratory facilities for a wide range dynamic fluorescence imaging, fabrication, and mechanobiophysics. Core D personnel have pro- fessional experience spanning the fields of high-resolution imaging and examining physical and chemical properties, including mechanobiophysics, and various scanning probe microscopies. All Core D experiments will be conducted at the University of Arizona in Tucson.