Live Cell Imaging and Protein Analytical Core

活细胞成像和蛋白质分析核心

• 批准号: 8616125
• 负责人: Ian Bird
• 金额: $ 11.02万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8616125
• 关键词: Antibodies; Calibration; Cell Communication; Cells; Cost Savings; Data Collection; Detection; Endothelial Cells; Ensure; Freezing; Human Resources; Image; Image Analysis; Life; Maintenance; Maternal-Fetal Exchange; Methodology; Methods; Microscopy; Procedures; Program Research Project Grants; Protein Analysis; Proteins; Reagent; Savings; Secure; Supporting Cell; Training; Training Support; Western Blotting; analytical method; assay development; base; cell preparation; cellular imaging; data acquisition; imaging modality; improved; instrument; method development; novel

In the context of preparing for this upcoming Program Project Grant (POl) renewal, over the last five years we have developed novel and cutting edge imaging methodology and established advanced protein analytical methods related to 0x43 analysis, as well as further training an expanding group of project personnel in general Ca2+ imaging and 0x43 protein analysis. We have also developed considerable expertise in dual Ca2+/N0 detection in endothelial cells of intact vessels and all these methods are now increasingly central to the projects of this combined POl. In the current application, continued training of current and new personnel and ongoing technical support of imaging methods application will continue through this new Core B and ongoing instrument technical support will be added for imaging microscopy. In addition, the Core will provide training support for cell protein analysis by Western blot based and Co-IP procedures as well as new antibody and technical assay development for all four projects. As a coordinating unit, the Core will offer further support to enhance data collection uniformity by relieving project personnel of the burden of preparing frozen stocks of validated pooled endothelial cells for imaging, so ensuring both high qualities of cell preparations providing uniformity of data acquisition between the projects. Where opportunities for reagent purchase can improve cost savings, or at least ensure more abundant reagents are fully used, the Core will coordinate those supplies to secure savings.

在为即将到来的计划项目拨款 (POl) 更新做准备的背景下，在过去五年中，我们开发了新颖且前沿的成像方法，建立了与 0x43 分析相关的先进蛋白质分析方法，并进一步培训了不断扩大的项目人员队伍，以进行一般 Ca2+ 成像和 0x43 蛋白质分析。我们还在完整血管内皮细胞的双重 Ca2+/N0 检测方面开发了相当多的专业知识，并且所有这些方法现在越来越成为该组合 POl 项目的核心。在当前的应用中，对现有人员和新人员的持续培训以及成像方法应用的持续技术支持将通过这个新的核心 B 继续进行，并且将为成像显微镜添加持续的仪器技术支持。此外，核心将为基于蛋白质印迹和 Co-IP 的细胞蛋白分析提供培训支持 所有四个项目的程序以及新抗体和技术测定的开发。作为协调单位，核心将提供进一步的支持，通过减轻项目人员准备用于成像的经过验证的混合内皮细胞冷冻储备的负担来提高数据收集的一致性，从而确保细胞制剂的高质量以及项目之间数据采集的一致性。如果试剂购买机会可以节省成本，或者至少确保充分利用更丰富的试剂，核心将协调这些供应以确保节省成本。