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.