用于脑组织解剖和固定的小鼠经心脏灌注
Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation.
作者信息
Wu Jinyun, Cai Yuqi, Wu Xiaoyang, Ying Yue, Tai Yilin, He Miao
机构信息
Institutes of Brain Sciences, Fudan University, Shanghai 200032, China.
出版信息
Bio Protoc. 2021 Mar 5;11(5):e3988. doi: 10.21769/BioProtoc.3988.
Abstract
Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and DNA molecules to preserve tissue and cell structure. Here we provide a step by step guide for performing this procedure in mouse.
摘要
先用生理盐水进行经心灌注，然后用4%多聚甲醛（PFA）灌注，这一方法被广泛用于清除血液并保存大脑，以进行免疫染色或杂交。PFA在溶液中分解为甲醛，甲醛可交联蛋白质和DNA分子以保存组织和细胞结构。在此，我们提供一份在小鼠中进行该操作的分步指南。
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本文引用的文献
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