Macromolecular Interaction Core

动脉粥样硬化和脂蛋白分析核心设施的范围是定量评估小鼠模型中两个位置的动脉粥样硬化病变面积。对于主动脉根试验，每位研究者仅负责提供在10%磷酸盐缓冲福尔马林中的离体盐水灌注心脏。然后，核心人员将：1）将心脏包埋在明胶中，2）将明胶块在OCT中冷冻，3）制备5个载玻片，每个载玻片包含4个12.5微米厚的切片，其覆盖主动脉根的前500微米，4）将载玻片用油红O、苏木精和快速绿色染色，以及5）定量一个载玻片上的病变面积。 每个载玻片的切片，并计算每个切片的平均病变。对于表面检测，核心将提供一名经过培训的技术人员，根据项目负责人批准的方案开展工作，以：1）麻醉小鼠，2）进行盐水灌注，通过放血使小鼠安乐死，3）解剖小鼠以原位显示小鼠主动脉，4）移除主动脉并修剪掉外膜组织，5）纵切主动脉并将其固定在两个显微镜载玻片之间，6）用油红O染色主动脉，7）将主动脉固定在显微镜上 幻灯片，以及8）量化整个主动脉中以及主动脉弓、胸主动脉和腹主动脉的区域中的表面损伤面积%。

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.