Jun 11, 2026
Cerebrospinal Fluid Collection (Terminal)
- Michael Henderson1,
- Lindsay Meyerdirk1,
- Libby Breton1
- 1Van Andel Institute
Protocol Citation: Michael Henderson, Lindsay Meyerdirk, Libby Breton 2026. Cerebrospinal Fluid Collection (Terminal). protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr1eybgmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 11, 2026
Last Modified: June 12, 2026
Protocol Integer ID: 318958
Keywords: csf from mice, cerebrospinal fluid collection, collecting csf, mice
Funders Acknowledgements:
ASAP
Grant ID: 020616
Abstract
Protocol for collecting CSF from mice.
Guidelines
All experiments involving animals must comply with federal and local law. All procedures should be reviewed and approved by the institutional animal care and use committee (IACUC) or equivalent ethics committee.
Materials
Anesthesia
Syringe
needle
Hair razor
Ethanol
Tape, tube rack, paper towel
Small microcentrifuge tubes (for collection)
Cotton swab sticks
Forceps
microscissors
Needle
20 µL pipet and gel loading tips
Safety warnings
BE CAREFUL TO AVOID BLOOD VESSELS!
Ethics statement
IACUC approved protocol 23-10-023.
Inject IACUC approved anesthesia to one mouse via intraperitoneal injection, and place mouse in bucket long enough for anesthesia to take effect. Apply a hard toe pinch until mouse no longer reacts, ensuring that the mouse can no longer feel pain before proceeding.
Shave the hair around the back of the mouse skull in area to be incised. Clean excess hair from surgery site with ethanol.
Place mouse on tube rack covered with paper towel, as indicated in image. Tape over mouse snout with tape with triangle cut out to allow mouse to breath.
Grasp skin on top of skull with forceps and cut longitudinally with small scissors. BE CAREFUL TO AVOID BLOOD VESSELS! As you are cutting, gently scrape apart the fascia that hold the skin to the muscle. Fold skin back to either side of incision line.
Grasp first layer of muscle with forceps and carefully cut down the middle and transversely where the muscle attaches to the skull. Ply apart from other muscle as you cut. Fold back muscle. BE CAREFUL TO AVOID BLOOD VESSELS!**
* If at some point you nick a blood vessel, apply water or ethanol to the cut and dry with cotton swab sticks until all blood and fluid has been cleared from surgery site.
Repeat the same procedure as in 5 with second and third layer of muscle. As you clear the last layer of muscle, be careful around the back of the skull. This is where there is only a thin membrane over the brain and where you will collect CSF.
Once area is cleared of muscle and fluid, grasp needle in one hand and 20 µL pipet with gel loading tip in the other hand. Puncture a hole with the needle and quickly place pipet tip over the hole. Slowly draw up CSF.
* If fluid is not coming, you may need to adjust the pipet tip location or puncture a larger hole. As you remove fluid, you should see the membrane drop towards the brain, indicating a removal of CSF. Extra CSF may be obtained by gently massaging mouse body.
Pipet CSF into small collection tube and spin down to remove contaminants. Freeze at -20°C.