microsphere embolic stroke rat
Microsphere embolic stroke in rats uses 20-50 micrometre spheres of materials such as dextran or superparamagnetic iron oxide to produce embolic occlusion and multifocal ischemic injury.
结局指标
存活脑组织染为红色、梗死区呈白色；以 2 mm 连续冠状切片逐孔染色成像后，测量患侧与对侧半球的梗死面积及总面积，计算患侧相对对侧的梗死面积百分比。
实验动物
- 年龄
- 成年（来源未报告具体周龄）
- 组织
- 脑组织（2 mm 冠状切片；梗死区与存活区）
- 年龄
- 成年（来源未报告具体周龄）
- 组织
- 脑组织（2 mm 冠状切片；梗死区与存活区）
实验步骤
TTC Staining TTC Staining The brain is removed from the skull and placed into a brain slicer mold placed in ice. Razor blades are positioned in the mold to obtain 2mm thick sections. Sections are placed in individual wells of a 24 well tissue culture dish with 500 μl of 2% TTC in 0.1M PBS. The plate is wrapped in foil and placed on a 37°C slide warmer for 30 minutes. TTC is removed and 500ul of fixative is added to each well for storage. Images of the sections are captured by Olympus S2H10 research stereo- microscope and Adobe Photoshop software. The infarct area and total area of the contralateral and ipsilateral hemispheres are measured with Scion image software (Scion Corporation). The percent infarct area is calculated as a percentage of the ipsilateral hemisphere relative to the contralateral hemisphere as previously described. [F2.JPG]]()
模型身份
- 模型名称
- microsphere embolic stroke rat
- 物种
- rat
生物学特性
- 表型出现阶段
- Embolic occlusion and infarct evolution are measured in the acute to subacute period; sphere size and dose determine lesion distribution.
- 适用研究问题
- Embolic stroke; microvascular occlusion; infarct variability; thrombolysis and reperfusion.
- 模型局限
- Microsphere material, size and dose vary between laboratories. It is not a blood-clot thromboembolic model and does not model fibrin-rich clot lysis; lesion distribution can be multifocal.
实验方法
实验方案
- 脑切片 TTC 染色与梗死面积测定
新鲜脑以2mm冠状切片逐孔经2% TTC（0.1M PBS）37°C避光染色30分钟后固定保存，体视成像并以患侧/对侧半球面积比计算梗死面积百分比。
本方案对该模型的适用性
状态：已验证适用· 证据类型：文献证据· 来源：PMID:26170628; https://pubmed.ncbi.nlm.nih.gov/26170628/
年龄：成年（来源未报告具体周龄）组织：脑组织（2 mm 冠状切片；梗死区与存活区）
大鼠微球栓塞后 2 mm 冠状脑切片 TTC 染色，定量栓塞性梗死范围。
- 脑切片 TTC 染色与梗死面积测定
新鲜脑以2mm冠状切片逐孔经2% TTC（0.1M PBS）37°C避光染色30分钟后固定保存，体视成像并以患侧/对侧半球面积比计算梗死面积百分比。
本方案对该模型的适用性
状态：已验证适用· 证据类型：文献证据· 来源：PMID:26170628; https://pubmed.ncbi.nlm.nih.gov/26170628/
年龄：成年（来源未报告具体周龄）组织：脑组织（2 mm 冠状切片；梗死区与存活区）
大鼠微球栓塞后 2 mm 冠状脑切片 TTC 染色，定量栓塞性梗死范围。
配套试剂与材料
- 2% TTC 染液
TTC 工作液（染色）
- 0.1M PBS缓冲液（TTC染色溶剂）
TTC 染色工作液的缓冲体系
- 组织固定液（TTC染色后保存用）
TTC 染色后的组织切片保存
- 组织固定液（TTC染色后保存用）
TTC 染色后的组织切片保存
- 0.1M PBS缓冲液（TTC染色溶剂）
TTC 染色工作液的缓冲体系
- 2% TTC 染液
TTC 工作液（染色）
参考文献
- Fluri F, Schuhmann MK, Kleinschnitz C. Animal models of ischemic stroke and their application in clinical research.. Drug design, development and therapy. 2015. PMID:26170628.
- Ian Simpson. Induce Stroke via Hypoxia-Ischemia. protocols.io, v2, 2019. DOI: 10.17504/protocols.io.8e5htg6
- Hu Y, Yang Z-H, Yan F, et al. CCA repair or ECA ligation - Which middle cerebral artery occlusion is better in the reperfusion mouse model? Ibrain. 2023;9(3):258-269. DOI:10.1002/ibra.12128; PMID:37786756.
- Bederson JB, Pitts LH, Germano SM, Nishimura MC, Davis RL, Bartkowski HM. Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats. Stroke. 1986;17(6):1304-1308. DOI:10.1161/01.STR.17.6.1304; PMID:2433817. 原文以成年大鼠 MCA 闭塞后 TTC 与 H&E 对比，24 小时 TTC 梗死面积 21±2% 的冠状切片面积，r=0.91，支持 TTC 用于大鼠脑梗死检测与定量。
- Clark WM, Lessov N, Lauten JD, Hazel K. Doxycycline treatment reduces ischemic brain damage in transient middle cerebral artery occlusion in the rat. J Mol Neurosci. 1997;9(2):103-108. DOI:10.1007/BF02736854; PMID:9407391. 原文在成年 Sprague-Dawley 大鼠以线栓行 2 小时可逆 MCAO，48 小时以梗死体积为终点（对照 158±44 mm3），支持 SD 大鼠 MCAO 梗死体积检测背景。
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