Sectioning and Counting of Motor Neurons in the L3 to L6 Region of the Adult Mouse Spinal Cord.
Sectioning and Counting of Motor Neurons in the L3 to L6 Region of the Adult Mouse Spinal Cord.
复制标题
成年小鼠脊髓 L3 至 L6 区域运动神经元的切片和计数。
DOI：
10.1002/cpz1.428
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Austin A
中科院分区：
文献类型：
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作者：
Austin A
组织学是研究组织的微观结构的学科。该方案允许产生腰髓区域L3至L6的冰冻横切面。它能够以一种可重复的方式对小鼠腹角腰运动神经元进行计数。方法包括脊柱解剖、液压挤压和组织学处理。冷冻切片的准备包括将腰髓埋入最佳切割温度(OCT)介质中。组织的正确方向在计算要丢弃的组织总量时至关重要。描述了有关截面厚度和安装的具体细节。这些要求不仅允许对特定区域的最佳覆盖，而且还确保没有单个运动神经元被计算两次。用半乳花青素对运动神经元的尼氏小体进行染色。所获得的截面都属于可比较的区域，质量保证是一致的。这种染色的特异性使这位科学家能够识别并可靠地量化腰部运动神经元。©2022作者。目前由Wiley期刊LLC发表的基本方案1：小鼠的安乐死和脊髓的分离基本方案2：脊髓的液压挤压基本方案3：腰区的识别基本方案4：在OCT中埋入脊髓基本方案5：收集冰冻切片到玻片上基本方案6：Gallocyanin染色基本方案7：运动神经元计数
Histology is the study of the microscopic structure of tissues. This protocol permits the generation of frozen transverse sections of lumbar spinal cord regions L3 to L6. It enables counting of murine ventral horn lumbar motor neurons in a reproducible manner. Methods include spinal column dissection, hydraulic extrusion, and histological processing. The preparation for cryo‐sectioning includes embedding lumbar spinal cord in optimal cutting temperature (OCT) medium. The correct orientation of the tissue is critical as calculating the amount of tissue to discard saved time overall. Specific details regarding section thickness and mounting are described. These requirements not only allow optimum coverage of specific regions but also ensure that no individual motor neuron was counted twice. The Nissl bodies of the motor neurons were stained using gallocyanin. The sections obtained are all of a comparable area and quality assurance is consistent. The specificity of the staining enables the scientist to identify and reliably quantify lumbar motor neurons. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Euthanasia of mouse and isolation of spinal cordBasic Protocol 2: Hydraulic extrusion of the spinal cordBasic Protocol 3: Identification of the lumbar regionBasic Protocol 4: Embedding cord in OCTBasic Protocol 5: Collection of frozen sections onto slidesBasic Protocol 6: Gallocyanin stainingBasic Protocol 7: Motor neuron counting