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小鼠模型两个位置的动脉粥样硬化病变区域的定量评估。对于主动脉根测定法，每个研究者仅负责在10％磷酸盐缓冲福尔马林中提供切除的盐水渗透的心脏。然后，核心人员将：1）将心脏嵌入明胶中，2）在10月3日冻结明胶块，3）准备5个包含四个12.5微米厚的部分，覆盖了主动脉根的前500微米，4）在油红色O，苏毒素和5）量子区域中染色。 每片截面并计算平均病变为每个部分。 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幻灯片，6）用油红O，7）将主动脉固定在显微镜上 幻灯片和8）量化整个主动脉以及主动脉弓的区域以及胸主动脉和腹主动脉的区域。