Anatomy-oriented stereotactic approach to cerebrospinal fluid collection in mice

Extraction of cerebrospinal fluid (CSF) in small animal models is an indispensable technique used in neuroscience and neuro-oncology research but can be technically challenging due to the small size of the brain. Here we describe a simple, reliable, and highly reproducible method for collecting CSF from anesthetized laboratory mice based on anatomical foundation, with no requirement of introducing any custom-made types of equipment. The mouse's head is fixed to a stereotaxic frame in the position that allows the maximum opening of the cisterna magna. A Hamilton syringe attached with a 26 gauge needle is inserted stereotactically in parallel to the brain axis to reach the cistern. The clear CSF (approximately 10 µl) can then be drawn slowly in approximately 5 min. Our modified approach minimizes technical variations that can result from customized equipment, and is applicable in broad research settings.