A step-by-step guide for microsurgical collection of uncontaminated cerebrospinal fluid from rat cisterna magna

Lennart Barthel; Harald Engler; Martin Hadamitzky; Laura Lückemann; Ulrich Sure; Manfred Schedlowski; Susann Hetze

doi:10.1016/j.jneumeth.2021.109085

A step-by-step guide for microsurgical collection of uncontaminated cerebrospinal fluid from rat cisterna magna

J Neurosci Methods. 2021 Mar 15:352:109085. doi: 10.1016/j.jneumeth.2021.109085. Epub 2021 Jan 27.

Authors

Lennart Barthel¹, Harald Engler², Martin Hadamitzky³, Laura Lückemann⁴, Ulrich Sure⁵, Manfred Schedlowski⁶, Susann Hetze⁷

Affiliations

¹ 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. Electronic address: [email protected].
² Institute of Medical Psychology and Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany. Electronic address: [email protected].
³ Institute of Medical Psychology and Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany. Electronic address: [email protected].
⁴ Institute of Medical Psychology and Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany. Electronic address: [email protected].
⁵ Department of Neurosurgery, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany. Electronic address: [email protected].
⁶ Institute of Medical Psychology and Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, 45147, Essen, Germany; Department of Clinical Neuroscience, Osher Center for Integrative Medicine, Karolinska Institutet, 171 77, Stockholm, Sweden. Electronic address: [email protected].
⁷ 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. Electronic address: [email protected].

PMID: 33508407
DOI: 10.1016/j.jneumeth.2021.109085

Abstract

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.

Keywords: CSF sampling; Central nervous system; Cerebrospinal fluid; Cisterna magna; Microsurgical technique; Rat; brain-immune communication pathways.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cerebrospinal Fluid
Cisterna Magna* / surgery
Punctures*
Rats
Specimen Handling