Immunohistochemistry

核心B：免疫组织化学（IHC）核心将提供组成和结构表征 血管病变（动脉粥样硬化、主动脉瘤和股动脉导丝诱导损伤） （项目1和4），以及血流动力学对血管基因表达的时空调控 力和高胆固醇血症（项目5）使用标准化的协议。服务包括（i）培训 个体项目中的人员，以适当地收获和包埋组织用于IHC分析，（ii）切割和 冷冻切片的安装方式，允许定量分析的形态区域， （iii）组织化学和免疫组织化学染色以及连续的 （iv）对处理过的组织进行图像分析，并对工作人员进行定量图像分析方面的培训， （五）编制出版物质量综合材料并进行培训， 图像.除了已经标准化的用于检测例如各种渗透的协议之外， 炎性细胞类型、泡沫细胞、内皮细胞、血管平滑肌细胞（在炎症的各个阶段）。 分化），循环细胞，凋亡细胞和细胞外基质成分，核心将发展 用于检测其他抗原（如血小板参与的PAF）的优化方案， 氧化反应和未折叠蛋白反应的标记物，以支持每个项目。的 核心总监将提前参与设计需要IHC分析的动物研究，以确保 来自每组最佳数量动物和最佳时间点的组织可用于分析， 并确定需要分析的适当目标组，以解决关于下列问题的具体假设： 病变的发展、组成和结构。核心导演 将帮助来自不同项目的个人解释IHC结果。核心活动将利用现有 设备，包括Microm HM 505 E低温恒温器、Leica Leitz DMRD直立显微镜（带明视野） 使用具有明场照明的Nikon E 600正置显微镜。Image Pro Plus图像分析软件（Media Cybernetics，银Spring，MD）、HP Color Laserjet 2550 LN和Canon Image Press Cl打印机。制定了协调时间安排和确保公平获得核心服务的程序。Core提供的组织化学和免疫组织化学分析将为量化病变程度提供额外的方法，并将为机制提供重要的见解 其调节病变的程度以及可影响结果的病变的性质，即，纤维化相对于炎性动脉粥样硬化病变更多，其可转化为斑块破裂的脆弱性。

Core B: The Immunohistochemstry (IHC) Core will provide characterization of the composition and architecture of vascular lesions (atherosclerosis, aortic aneurysms and femoral artery wire-induced injury) (Projects 1 & 4), and ofthe temporal and spatial regulation of vascular gene expression by hemodynamic forces and hypercholesterolemia (Project 5) using standardized protocols. Services include (i) training personnel in individual projects to harvest and embed tissue appropriately for IHC analysis, (ii) cutting and mounting of cryostat sections in a manner that allows quantitative analysis of morphologic regions over defined areas of lesion, (iii) histochemical and immunohistochemical staining and counterstaining of serial sections, (iv) image analysis of processed tissue and training in quantitative image analysis of personnel in individual projects, (v) generation of and training in the generation of publication quality composites of images. In addition to already standardized protocols for the detection of for example, various infiltrating inflammatory cell types, foam cells, endothelial cells, vascular smooth muscle cells (at various stages of differentiation), cycling cells, apoptotic cells and extracellular matrix components, the core will develop optimized protocols for detection of additional antigens (such as PAF for platelet involvement), markers of oxidative responses and markers of the unfolded protein response as needed to support each project. The Core Director will participate up front in designing animal studies that will require IHC analysis to ensure that tissues from optimal numbers of animals per group and at optimal time points will be available for analysis, and in determining the appropriate panel of targets to be analysed to address specific hypotheses regarding mechanisms underlying lesion the development, composition and architecture of lesions. The Core Director will assist individuals from the various projects in interpreting IHC results. Core activities will utilize existing equipment including a Microm HM505 E cryostat, Leica Leitz DMRD upright microscope with bright field illumination, Nikon E 600 upright microscope with bright field illumination. Image Pro Plus image analysis software (Media Cybernetics, Silver Spring, MD), HP color laserjet 2550 LN and Canon Image Press Cl printers. Procedures for coordinated scheduling and for assuring equitable access to core services are in place. The histochemical and immunohistochemical analysis provided by the Core will provide additional approaches for quantification of the extent of lesion and will provide important insights into the mechanisms that modulate the extent of lesions as well as the properties of lesions that can impact outcome, i.e. more fibrotic versus inflammatory atherosclerotic lesions that may translate into vulnerability to plaque rupture.