LDL receptor-deficient rabbit
LDL 受体缺失兔在普通或高胆固醇饲料下出现高胆固醇血症和主动脉粥样硬化病变,可用于研究 LDL 清除障碍驱动的病变。
结局指标
获得可用于兔主动脉斑块组织学分级和组成的 HE 染色切片。
实验动物
- 年龄
- not reported in the cited source
- 组织
- aortic root; thoracic aorta
实验步骤
H&E staining **Dewaxing:** Paraffin sections are placed in xylene for 3 x 00:03:00 H&E staining **Rehydration: ** Paraffin sections are placed in - 100% EtOH for 3 x 00:03:00 - 95% EtOH for 1 x 00:03:00 - Rinse with water H&E staining **Hematoxylin staining: ** Paraffin sections are stained with hematoxylin for 00:00:20 and rinse under running water for 00:01:00 H&E staining **Differentiation and bluing:** Paraffin sections are placed into the bluing reagent for 00:01:00 and rinse under running water for 00:01:00 _This step results in a blue coloration of the nuclei._ H&E staining **Eosin staining:** Parrafin sections are counterstained with Eosin for 00:00:10 _This step results in a pink coloration of the cytoplasmic components and connective tissue fibers._ H&E staining **Dehydration and Clearing:** Paraffin sections are placed in - 95% EtOH for 00:02:00 - 100% EtOH for 2 x 00:02:00 - Xylene for 2 x 00:02:00 H&E staining **Mounting:** Parrafin sections are mounted with a glass coverslip using Permount.
模型身份
- 模型名称
- LDL receptor-deficient rabbit
- 物种
- rabbit
- 基因型/合子状态
- Ldlr -/-
- 等位基因
- Ldlr
生物学特性
- 表型出现阶段
- 高胆固醇饲料下病变明显;普通饲料下进展取决于遗传背景和年龄。
- 适用研究问题
- LDL 受体缺失兔的脂蛋白代谢和主动脉病变;基因编辑兔用于动脉粥样硬化研究的适用性。
- 模型局限
- 基因编辑兔的品系和突变细节需按具体来源核对;兔模型成本高、繁殖周期长;病变仍不能完全复制人类斑块破裂和血栓事件。
实验方法
实验方案
- 动脉组织石蜡切片 H&E 染色
通过苏木素与伊红显示动脉组织的细胞核、细胞质及结缔组织结构。
配套试剂与材料
- Epredia hematoxylin 7221
细胞核染色
- Fisherbrand xylene HC7001GAL
脱蜡或透明
- Fisherbrand xylene HC7001GAL
脱蜡或透明
- Ethanol working reagent, 100%
复水或脱水
- Ethanol working solution, 95%
复水或脱水
- Ethanol working reagent, 100%
复水或脱水
- Ethanol working solution, 95%
复水或脱水
- Epredia bluing reagent 7301
返蓝
- Epredia eosin Y 7111
细胞质及结缔组织复染
- Fisher Permount mounting medium SP15
封片
- Water for histological staining and rinsing
染色冲洗
- Water for histological staining and rinsing
染色冲洗
- Water for histological staining and rinsing
细胞核染色
参考文献
- Fan J, et al. Rabbit models for the study of human atherosclerosis: from pathophysiological mechanisms to translational medicine. Pharmacology & Therapeutics. 2015;146:104-119. DOI:10.1016/j.pharmthera.2014.09.009; PMID:25277507.
- protocols.io · vv1 · Nathalia L. Oliveira; samantha.gruenheid · This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
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