Post-mortem Examination of Animals
Post-mortem Examination of Animals
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DOI：
10.1007/978-2-8178-0472-9_7
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Vogt, Catherine
中科院分区：
文献类型：
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作者：
Ferry, Barbara;Gervasoni, Damien;Vogt, Catherine
在使用立体定位手术来操纵特定大脑目标结构的活动的实验结束时，有必要对大脑进行尸检以验证实验数据。本章对安乐死、尸检和脑切除、器官固定、切片和染色等不同步骤进行了全面的描述，安乐死的方法必须与实验相适应，并确保动物的人道死亡。麻醉药物过量是合适的，当大脑在被移除之前必须用固定剂灌流。当动物在适当的止痛下被深度麻醉时，可以在心脏仍在跳动的情况下通过灌流对大脑进行固定。一旦固定，大脑就会被取出，准备切片。冰冻的大脑切片是使用冰冻恒温器制作的，根据研究的目的，冰冻切片的厚度各不相同。然后将切片安装在冷凝玻璃载玻片上，在室温下烘干24小时，或在37°C的烤箱中烘干12小时。染色后，在光学显微镜下检查每个切片。在对所有切片进行有条不紊的审查之后，在组织学上确认靶点正确位置的动物可以被包括在统计分析中。提供操纵地点的证据允许对设计和执行实验的坐标进行核实。只有在这种验证之后，才能在采集的数据和目标站点之间进行关联。
At the end of an experiment in which a stereotaxic surgery has been used to manipulate the activity of a particular brain target structure, postmortem examination of the brain is necessary to validate the experimental data. The different steps including euthanasia, autopsy and removal of the brain, treatments specific to organ fixation, sectioning, and staining are comprehensively described in this chapter.The methods of euthanasia must be compatible with the experiment and ensure a humane death of the animal. Anesthetic overdose is suitable when the brain has to be perfused with a fixative before being removed. When the animal is deeply anesthetized with appropriate analgesia, the fixation of the brain by perfusion can be performed while the heart is still beating. Once fixed the brain can is removed and prepared for sectioning. Frozen brain sections are made using a cryostat with various thicknesses depending on the purpose of the study. Sections are then mounted on cold gelatinized glass slides and dried at room temperature for 24 h or in an oven at 37 °C for 12 h. After staining, each section is examined under an optical microscope. Following a methodical review of all the sections, animals for which a correct location of the target is histologically confirmed can be included for statistical analyses. Providing evidence for the site of manipulation permits a verification of the coordinates under which the experiments were designed and performed. It is only after this verification that correlations between acquired data and target site can be made.