Biophysical Imaging

生物物理成像

• 批准号: 10871780
• 负责人: Carol C Gregorio
• 金额: $ 19.96万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-20 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10871780
• 关键词: 3-Dimensional; ABL1 gene; Acute Respiratory Distress Syndrome; Address; Arizona; Atomic Force Microscopy; Biological; Biophysics; Cells; Chicago; Complex; Computer software; Consultations; Core Facility; Correlative Study; Cytoskeleton; DOCK1 gene; Data; Data Analyses; Data Set; Directories; Doctor of Philosophy; EMS1 gene; Endothelial Cells; Endothelium; Engineering; Equipment; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Focal Adhesions; Genetic; Genomics; Health Insurance Portability and Accountability Act; Human; Human Resources; Image; Individual; Integrin beta4; Intercellular Junctions; Laboratories; Laboratory Research; Lasers; Lung; MYLK gene; Measurement; Measures; Mechanics; Membrane; Microfabrication; Microscopy; Molecular; Mutation; Names; Optics; Organelles; P-Selectin; Post-Translational Protein Processing; Procedures; Process; Proteome; Quantitative Microscopy; Regulation; Research Personnel; Research Project Grants; Resolution; Resources; Retrieval; Sampling; Scanning; Scanning Probe Microscopes; Scanning Probe Microscopy; Scientist; Secure; Services; Signal Transduction; Site; Slide; Standardization; Structure; System; TLR4 gene; Technology; Text; Time; Traction Force Microscopy; Training; Transgenes; Universities; University resources; biophysical analysis; cellular engineering; chemical property; encryption; experience; experimental study; fabrication; fluorescence imaging; high resolution imaging; imaging study; instrument; laboratory facility; live cell imaging; multimodality; nano; paxillin; physical property; receptor; superresolution microscopy; three dimensional structure; tissue culture; tool; web portal

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.

摘要： 生物物理成像核心的主要目标是为PPG研究者提供完整的范围 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 与内皮细胞（EC）细胞骨架重排和屏障的细胞和分子基础有关 调控核心D人员和设备将使PPG科学家能够对3D结构进行成像，评估物理结构， 校准和化学性质，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junc， 选择。该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，总内部 反射荧光（TIRF）显微镜，动态（现场）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显微镜， 显微镜以允许观察细胞器（例如，皮质细胞骨架，连接复合体，局灶性附加， hesions和lamellipodia）的详细信息（3D）。核心D还将与每个项目和核心B（Ge- nomic/GeneticandProteomeCore）来培养经工程改造以表达不同nmMLCK 1/2的内皮细胞， coronin、NAMPT、TLR4、DOCK 1、lamellipodin、integrin β4、kindlin-2、PSGL 1、P-Selectin和S1PR 1/3 recep。 检测具有特定SNP或翻译后修饰突变的转基因，并测量各种生物物理 力（TFM和AFM）。核心还提供访问经验丰富的使用的完整资源的大学， 亚利桑那大学成像核心设施，包括最先进的高分辨率荧光显微镜 系统.该核心还将利用亚利桑那州研究实验室的公共资源。 生物物理成像核心，由卡罗尔C。格雷戈里奥，博士，包括人员和实验室设施， 宽范围动态荧光成像、制造和机械生物物理学。核心D人员有亲- 专业经验跨越高分辨率成像和检查物理和化学领域 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。