大鼠枕大池无污染脑脊髓液的显微采集分步指南。
A step-by-step guide for microsurgical collection of uncontaminated cerebrospinal fluid from rat cisterna magna.
机构信息
Department of Neurosurgery, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany; Institute of Medical Psychology and Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany.
Institute of Medical Psychology and Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany.
出版信息
J Neurosci Methods. 2021 Mar 15;352:109085. doi: 10.1016/j.jneumeth.2021.109085. Epub 2021 Jan 27.
BACKGROUND
Cerebrospinal fluid (CSF) analysis is of significant clinical importance for the diagnosis of diseases. In humans, CSF is easily accessible and can be collected using minimally invasive methods. However, obtaining uncontaminated CSF from rats is still challenging.
NEW METHOD
This study described a microsurgical technique for sampling large quantities (>200 μL) of clear and non-blood-contaminated CSF from the rat cisterna magna in a comprehensible step-by-step guide and provided a graphical visualization.
RESULTS
CSF was sampled in 5-10 min (n = 29 animals; average surgical time 7.6 min). In visual control, 28 samples (97 %) of clear and uncontaminated CSF were obtained. The volume of CSF collected was 124-337 μL, with an average volume of 207 μL/sample. Using the Valsalva maneuver, we could collect higher volumes (up to 400 μL) several times.
COMPARISON WITH EXISTING METHOD(S): There is no need for special surgical skills to perform this method accurately. The method takes a few minutes longer than a percutaneous puncture (<1 min in pups). However, the volume of CSF obtained using the percutaneous approach in adult rats (50-70 μL) is comparatively low.
CONCLUSIONS
We described a practical method of sampling CSF from rats that enables large volumes of CSF to be collected without blood contamination. No special surgical skills are required to use this method. With proper practice, the time between skin incision and CSF sampling is <10 min. Depending on the experimental design requirements, some additional time must be planned for wound closure.
背景
脑脊液（CSF）分析对于疾病的诊断具有重要的临床意义。在人类中，CSF 很容易获得，可以通过微创方法收集。然而，从大鼠中获得无污染的 CSF 仍然具有挑战性。
新方法
本研究描述了一种从大鼠枕大池获取大量（>200 μL）清亮、无污染 CSF 的显微外科技术，提供了通俗易懂的分步指南和图形可视化。
结果
CSF 在 5-10 分钟内（n = 29 只动物；平均手术时间 7.6 分钟）被取样。在视觉控制下，获得了 28 份（97%）清亮、无污染的 CSF 样本。收集的 CSF 体积为 124-337 μL，平均每个样本为 207 μL。使用瓦氏动作，我们可以多次收集更高的体积（高达 400 μL）。
与现有方法的比较
执行此方法不需要特殊的手术技能。该方法比经皮穿刺术（<1 分钟）需要多几分钟时间。然而，在成年大鼠中经皮穿刺术获得的 CSF 体积（50-70 μL）相对较低。
结论
我们描述了一种从大鼠中采集 CSF 的实用方法，该方法可以在不污染血液的情况下采集大量 CSF。使用这种方法不需要特殊的手术技能。经过适当的练习，从皮肤切口到 CSF 取样的时间<10 分钟。根据实验设计要求，还需要计划一些额外的时间来关闭伤口。
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