小鼠大脑中动脉局部凝血酶血栓模型
脑动脉内局部凝血酶作用形成血栓,产生局灶性缺血损伤。
Outcome measures
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。
Experimental animals
- Tissue
- brain sections
Experimental procedures
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]]()
Model identity
- Model name
- 小鼠大脑中动脉局部凝血酶血栓模型
- Species
- mouse
Biological characteristics
- Research questions
- 缺血性脑损伤与模型适用性比较。
- Limitations
- 局部形成血栓与预制血凝块或微球栓塞的构建不同,不能以名称相近视为同一模型。
Experimental methods
Experimental protocols
- 脑切片 TTC 染色与梗死面积测定
Fresh brains are cut to 2-mm coronal sections, stained per well with 2% TTC in 0.1M PBS at 37°C for 30 min protected from light, stored in fixative, imaged on a stereomicroscope, and infarct percentage computed ipsilateral-versus-contralateral.
Reagents and materials
- 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)
References
- Preclinical models of middle cerebral artery occlusion: new imaging approaches to a classic technique.. PMID:37409019
- 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。
- ARRIVE 2.0 guidelines