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Integrated Protocol for Assessing Meningeal Lymphatics: Cisterna Magna Injection, Lymph Node Imaging, Meningeal
Baokai Zhang1, Rong Wang1, Shengli Zhao1
1School of Medicine, School of Life Sciences, Shanghai University.
Journal of Visualized Experiments : Jove
|June 15, 2026
Summary
This study presents a rapid 3-hour protocol to assess meningeal lymphatic vessels (MLVs) in mice. The method combines tracer injection, live imaging, and tissue analysis to evaluate MLV morphology and function for CNS homeostasis research.
Area of Science:
- Neuroscience
- Immunology
- Vascular Biology
Background:
- Meningeal lymphatic vessels (MLVs) are crucial for central nervous system (CNS) homeostasis.
- MLV dysfunction is linked to neurological disorders, necessitating robust evaluation methods.
Purpose of the Study:
- To establish an integrated protocol for assessing MLV morphology and function in mice.
- To provide a comprehensive method for CNS lymphatic system research.
Main Methods:
- Intra-cisterna-magna (i.c.m.) injection of fluorescent tracers into the cerebrospinal fluid (CSF).
- Live imaging of tracer drainage to cervical lymph nodes (CLNs).
- Meningeal dissection and whole-mount staining for dorsal MLV analysis.
Main Results:
- The protocol allows real-time visualization of tracer transport from CSF to CLNs.
- Detailed morphological analysis of dorsal MLVs is achievable through whole-mount staining.
- The entire procedure, including i.c.m. injection and tissue preparation, is completed within 3 hours.
Conclusions:
- This integrated protocol offers an efficient method for evaluating MLVs in mice.
- The protocol is adaptable for intracranial drug delivery and molecular analyses using fresh tissues.
