An objective procedure for ischemic area evaluation of the stroke intraluminal thread model in the mouse and rat
An objective procedure for ischemic area evaluation of the stroke intraluminal thread model in the mouse and rat
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DOI：
10.1016/s0165-0270(01)00476-9
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发表时间：
2002-01-15
影响因子：
3
通讯作者：
Kerr, JS
中科院分区：
文献类型：
--
作者：
Wexler, EJ;Peters, EE;Kerr, JS
计算机辅助程序用于测量动物中风模型中的梗死面积，但这种方法通常遵循不太客观的梗死边界的手动描记。建立在先前报道的方法，使用扫描图像的三苯基四唑氯化物（TTC）染色的大鼠脑内的管腔内线程模型，我们开发了一种客观的方法来评估缺血性损伤的小鼠和大鼠的大脑。我们的方法的独特之处在于使用假处理的动物，从而允许从同侧受伤的大脑中去除正常的大脑白色物质。使用Microtek Scanmaker 4平板扫描仪同时扫描每只动物的所有脑切片。对2 mm冠状脑切片的秋季彩色图像进行彩色分割。使用Image Pro Plus（4.0）和彩色分割，在绿色通道中测量缺血和正常白色物质区域，在红色通道中测量整个脑区域。计算假处理动物和脑缺血动物的未染色组织百分比。通过从缺血组的平均未染色面积中减去假处理组的平均未染色面积，计算缺血面积。使用小鼠和大鼠永久性和短暂性局灶性缺血模型以及永久性缺血模型中的MK-801对该方法进行了验证。在损伤前以3 mg/kg i. p.剂量给予MK-801，可使小鼠和大鼠永久性闭塞模型中的损伤分别减少75%和44%。这种方法的好处包括：客观的分析缺血性损伤，使用现成的软件，使成本可以控制和删除正常皮质下白色物质的计算。这种方法应该允许更一致的评价梗死面积的变化，因此，减少变异性和更高的生产力。(C)2002年由Elsevier Science B. V.出版，保留所有权利。
Computer-assisted procedures are used to measure infarct areas in animal stroke models, but this approach usually follows the less objective manual tracing of the boundaries of the infarct. Building on previously reported methodology using scanned images of triphenyltetrazolium chloride (TTC)-stained rat brains in the intraluminal thread model, we developed an objective method to assess ischemic damage in both the mouse and rat brains. The unique addition to our approach is the use of sham-treated animals, which thereby permits the removal of normal brain white matter from the ipsilateral injured brain. All brain sections per animal were scanned simultaneously using a Microtek Scanmaker 4 flatbed scanner. Color segmentation on fall color images of 2 mm coronal brain sections was performed. Using Image Pro Plus (4.0) and color segmentation, ischemic and normal white matter areas were measured in the green channel and the entire brain area in the red channel. The percent of unstained tissue was calculated for sham-treated animals and for those with cerebral ischemia. By subtracting the average unstained area of the sham-treated group from the average unstained area from the ischemic group, the ischemic area was calculated. This methodology was validated using mouse and rat permanent and transient, focal ischemia models and MK-801 in the permanent ischemia models. MK-801, dosed at 3 mg/kg i.p. prior to the injury, reduced the injury by 75% in the mouse and 44% in the rat permanent occlusion models. The benefits of this methodology include: objectivity of the analysis of the ischemic injury, use of readily available software so that costs can be contained and removal of normal subcortical white matter from the calculation. This method should allow more consistent evaluation of changes in the infarct size, therefore, resulting in reduced variability and higher productivity. (C) 2002 Published by Elsevier Science B.V. All rights reserved.