Whole animal perfusion fixation for rodents.
Whole animal perfusion fixation for rodents.
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DOI：
10.3791/3564
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发表时间：
2012-07-30
期刊：
影响因子：
--
通讯作者：
Shain, William
中科院分区：
文献类型：
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作者：
Gage, Gregory J;Kipke, Daryl R;Shain, William
固定的目标是快速、均匀地将组织保持在类似生命的状态。虽然将组织直接放置在固定剂中对于小块组织效果良好，但较大的标本（如完整的大脑）会给浸泡固定带来问题，因为固定剂不会以相同的速度到达组织的所有区域（5，7）。通常，在组织可以被保存之前，对缺氧的反应变化开始（12）。通过循环系统直接灌注固定剂的优点是，化学品可以利用天然血管网络快速到达生物体的每个角落。为了最有效地利用循环系统，必须注意与生理压力相匹配（3）。重要的是要注意，生理压力取决于所用的物种。灌注固定的技术根据待固定的组织以及固定后如何处理组织而变化。在本视频中，我们描述了一种低成本、快速、可控和统一的固定程序，使用4%多聚甲醛通过血管系统灌注：通过大鼠心脏，以获得最佳的大脑保存，用于免疫组织化学。这种技术的主要优点（相对于重力进料系统）是循环系统得到最有效的利用。
The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. While placing tissue directly in fixative works well for small pieces of tissue, larger specimens like the intact brain pose a problem for immersion fixation because the fixative does not reach all regions of the tissue at the same rate (5,7). Often, changes in response to hypoxia begin before the tissue can be preserved (12). The advantage of directly perfusing fixative through the circulatory system is that the chemical can quickly reach every corner of the organism using the natural vascular network. In order to utilize the circulatory system most effectively, care must be taken to match physiological pressures (3). It is important to note that physiological pressures are dependent on the species used. Techniques for perfusion fixation vary depending on the tissue to be fixed and how the tissue will be processed following fixation. In this video, we describe a low-cost, rapid, controlled and uniform fixation procedure using 4% paraformaldehyde perfused via the vascular system: through the heart of the rat to obtain the best possible preservation of the brain for immunohistochemistry. The main advantage of this technique (vs. gravity-fed systems) is that the circulatory system is utilized most effectively.