Live Cell Imaging and Protein Analytical Core

在为即将到来的计划项目资助（POI）更新做准备的背景下，在过去的五年里，我们开发了新的和尖端的成像方法，建立了与0x 43分析相关的先进蛋白质分析方法，并进一步培训了一批不断扩大的项目人员，包括一般的Ca 2+成像和0x 43蛋白质分析。我们还在完整血管内皮细胞的双重Ca 2 +/N 0检测方面积累了大量专业知识，所有这些方法现在都越来越成为该联合POL项目的核心。在当前应用中，将通过新的核心B继续对现有和新员工进行持续培训，并继续提供成像方法应用的技术支持，并将为成像显微镜增加持续的仪器技术支持。此外，该中心还将为基于蛋白质印迹和Co-IP的细胞蛋白质分析提供培训支持 为所有四个项目开发新的抗体和技术分析方法。作为一个协调单位，核心将提供进一步的支持，以提高数据收集的一致性，减轻项目人员准备经验证的合并内皮细胞冷冻储备用于成像的负担，从而确保细胞制备的高质量，提供项目之间数据采集的一致性。如果试剂采购的机会可以提高成本节约，或至少确保更丰富的试剂得到充分利用，核心将协调这些供应，以确保节约。

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.