Macromolecular Interaction Core

高分子相互作用核心

• 批准号: 8242738
• 负责人: PAUL L FOX
• 金额: $ 26.11万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8242738
• 关键词: Adventitious Tissue; Aorta; Area; Atherosclerosis; Biological Assay; Blood Vessels; Cells; Collaborations; Core Facility; Data; Fast Green; Formalin; Freezing; Gelatin; Green S; Heart; Hematoxylin; Human Resources; In Situ; Individual; Inflammatory Response; Laboratories; Learning; Lesion; Lipids; Lipoproteins; Location; Microscope; Mus; Perfusion; Phosphate Buffer; Plant Roots; Preparation; Protocols documentation; Research Personnel; Saline; Slide; Staining method; Stains; Standardization; Surface; Thick; Thoracic aorta; Tissues; Training; Transgenic Mice; Work; abdominal aorta; aortic arch; investigator training; mouse model; oil red O

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.

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病变区域进行定量评估。对于主动脉根测定，每个研究者仅负责提供在 10% 磷酸盐缓冲福尔马林中灌注生理盐水的切除心脏。然后，核心人员将：1) 将心脏嵌入明胶中，2) 在 OCT 中冷冻明胶块，3) 准备 5 个载玻片，每个载玻片包含四个 12.5 微米厚的切片，覆盖主动脉根部的前 500 微米，4) 染色油红 O、苏木精和固绿载玻片，以及 5) 对一张载玻片上的病变区域进行定量 每张幻灯片的切片并计算每个切片的平均病变。对于正面检测，核心将提供一名训练有素的技术人员，按照项目负责人批准的方案工作：1) 麻醉小鼠，2) 进行盐水灌注，通过放血使小鼠安乐死，3) 解剖小鼠以显示小鼠主动脉原位，4）取出主动脉并修剪外膜组织，5）纵向切割主动脉并将其压平固定在两个显微镜之间载玻片，6) 用油红 O 染色主动脉，7) 将主动脉安装在显微镜上 幻灯片，8) 量化整个主动脉以及主动脉弓、胸主动脉和腹主动脉区域的表面病变面积百分比。