Macromolecular Interaction Core

动脉粥样硬化和脂蛋白分析核心设施的范围是对小鼠模型中两个位置的动脉粥样硬化损伤区域进行定量评估。对于主动脉根部分析，每个单独的研究者只负责提供10%磷酸盐缓冲福尔马林中的切除的生理盐水灌流心脏。然后，核心人员将：1)将心脏包埋在明胶中，2)在10月份冷冻明胶块，3)准备5张幻灯片，每张包含4个12.5微米厚的切片，覆盖主动脉根部的前500微米，4)用油红O、苏木素和坚固绿色染色切片，5)在一张切片上定量显示病变区域 每张玻片切片，并计算每一切片的平均损伤数。对于EN Face测试，Core将提供一名训练有素的技术专家，根据项目负责人批准的方案进行以下工作：1)麻醉小鼠，2)进行生理盐水灌流，通过放血安乐死小鼠，3)解剖小鼠以原位显示小鼠的主动脉，4)切除大动脉并切除外膜组织，5)纵向切开大动脉并将其固定在两张显微镜载玻片之间，6)用油红O染色大鼠的主动脉，7)在显微镜下安装大动脉 幻灯片，和8)量化的百分比表面病变面积在整个主动脉，以及在区域的主动脉弓，胸腹主动脉。

The scope of the Atherosclerosis and Lipoprotein Analysis Core facility is the quantitative assessment of atherosclerosis lesion areas at two locations in mouse models. For the aortic root assay, each individual investigator is only responsible for providing the excised saline-perfused heart in 10% phosphate buffered formalin. The Core personnel will then: 1) embed the heart in gelatin, 2) freeze the gelatin block in OCT, 3) prepare 5 slides containing four 12.5 micron thick sections each, that cover the first 500 microns of the aortic root, 4) stain the slides in oil red O, hematoxylin, and fast green, and 5) quantitate the lesion areas on one section per slide and calculate the mean lesion are per section. For the en face assay, the Core will provide a trained technologist to work under the project leader's approved protocol to: 1) anesthetize the mice, 2) perform saline perfusion, which euthanizes the mouse by exsanguination, 3) dissect the mouse to display the mouse aorta in situ, 4) remove the aorta and trim off adventitial tissue, 5) cut the aorta longitudinally and fix it flattened between two microscope slides, 6) stain the aorta with oil red O, 7) mount the aorta on a microscope slide, and 8) quantify the % surface lesion area in the entire aorta, as well as in the regions of the aortic arch, and thoracic and abdominal aortas.