Department of Psychiatry and Behavioral Neurosciences, McMaster University, Psychology Building Rm 303, 1280 Main St. West, Hamilton, Ontario, 8S 4K1, Canada.
J Neurosci Methods. 2019 Jan 1;311:402-407. doi: 10.1016/j.jneumeth.2018.09.025. Epub 2018 Sep 28.
The composition of cerebrospinal fluid (CSF) is an invaluable parameter in better understanding of cellular and molecular processes within the mammalian brain. However, the collection of significant volumes of clean CSF can be technically challenging in studies with laboratory mice. Over the past five decades, several approaches have been developed to maximize the quantity and quality of CSF samples, either from live or euthanized animals. Due to the small amounts collected, samples from single mice were often pooled or diluted to meet volume requirements of automated counters and multiple assays.
This paper reviews previous work on CSF collection in mice, thus providing methodological background for the current post-mortem procedure. This modified cisternal puncture method involves the use of a peristaltic pump for consistent and slow intracardiac perfusion, as well as a loupe headset with a custom-made glass pipette for piercing a single hole in the atlanto-occipital membrane during repeated CSF draws. Sample cleanness is verified by comparing the colour of the glass pipette and the bottom of centrifuged PCR vial against a white background.
With three trained experimenters, the entire procedure (including anesthesia) takes ∼11-13 min and often results in the collection of up to 40 μl of clean CSF from males of different murine strains. Properly staggered collections allow processing of relatively large cohorts of mice per day.
This modification of previously employed methods can be used in studies that require tightly-timed collections of larger volumes of undiluted, tissue-free CSF and/or individual data records.
脑脊液（CSF）的成分是更好地了解哺乳动物大脑内细胞和分子过程的一个非常有价值的参数。然而，在实验室小鼠的研究中，收集大量清洁的 CSF 样本在技术上具有挑战性。在过去的五十年中，已经开发出几种方法来最大限度地增加 CSF 样本的数量和质量，无论是来自活体还是安乐死的动物。由于收集到的样本量很少，来自单个小鼠的样本通常会被汇集或稀释，以满足自动计数器和多个检测的体积要求。
本文回顾了以前在小鼠中 CSF 收集的工作，从而为当前死后程序提供了方法学背景。这种改良的脑池穿刺方法涉及使用蠕动泵进行一致和缓慢的心内灌注，以及带有定制玻璃移液管的体视镜耳机，用于在重复 CSF 抽取过程中刺穿枕骨寰枢膜上的单个孔。通过将玻璃移液管和离心 PCR 管底部与白色背景进行比较，来验证样本的清洁度。
通过三名训练有素的实验员，整个过程（包括麻醉）大约需要 11-13 分钟，通常可以从不同品系的雄性小鼠中收集多达 40 μl 的清洁 CSF。适当交错的采集允许每天处理相对较大的小鼠队列。
对先前使用的方法进行修改，可以用于需要严格时间采集较大体积的未稀释、无组织 CSF 和/或单个数据记录的研究。
