Jan 22, 2026
Stereotaxic surgery
- Oscar Moreno Ramos1,
- Divya Raj1
- 1Northwestern University
Protocol Citation: Oscar Moreno Ramos, Divya Raj 2026. Stereotaxic surgery. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvj1n3xvk5/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: January 22, 2026
Last Modified: January 23, 2026
Protocol Integer ID: 239187
Keywords: ASAPCRN, stereotaxic surgery stereotaxic surgery, stereotaxic surgery
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-020600
Abstract
Stereotaxic surgery
Materials
O2 tank,sterile instruments, drugs: isoflurane, buprenex,
meloxicam, 70% ethanol, Betadine, warming pad for stereotax and for recovery, eye
moistener, lidocaine for ears, solution to inject, syringes, wound clips, scalpel
Troubleshooting
Prior to surgery
Transfer cages to conventional or containment
Autoclave surgical instruments
Prepare everything needed for surgery (sterile instruments, drugs: isoflurane, buprenex, meloxicam, 70% ethanol, Betadine, warming pad for stereotax and for recovery, eye moistener, lidocaine for ears, solution to inject, syringes, wound clips, scalpel)
Pre-op
Ensure isoflurane reservoir is filled between the two black lines.
Open O2 tank
Adjust O2 flow rate to 0.8-1.5
Put mice in anesthesia chamber
Ensure line to stereotaxic apparatus is pinched
Set isoflurane to 3%
Once the mouse is anesthetized switch flow line to mask and set isofluorane to 1.5-2 and O2 rate to 0.8-1.5.
Inject mice with Buprenex subcutaneously at 0.1mg/kg; Buprenex, buprenorphine hydrochloride, comes in 0.3mg/mL solution. (a 30g mouse would be injected with 0.1mL)
Surgery
Unpinch isoflurane line to the stereotaxic apparatus, pinch line toward chamber
Install animal in the stereotaxic apparatus, use the mouthpiece and adjust the mask
Set isoflurane concentration to 1.8-2% and O2 rate to 0.5-0.8
Put eye moistener and ensure there’s plenty on the eye throughout surgery
Once the animal is anesthetized, secure the head in place with the ear pieces
Shave head
Prepare head with Betadine, followed by 70% ethanol
Ensure deep anesthesia by pinching one foot
Make a 5 mm incision in a rostro-caudal direction in between the ears
Expose the skull via blunt dissection of the overlying musculature and scrape off connective tissue
Identify landmark point bregma and calculate stereotaxic coordinates
Use the microdrill to puncture a small hole in the skull at the rostro-caudal/medio-lateral stereotaxic coordinates
Lower needle to the dorso-ventral coordinate over 30s to 1min (can add 0.02mm to create a pocket, then retract to the calculated z coordinate)
For viral vector solution, inject 0.2-0.4uL over a period of 5min (0.08uL/min), for cell solutions inject 1 to 2 ul over 5 min.
Wait 5 min after the injection, then withdraw the needle slowly
Close the skin with wound clips (make sure they’re not close to the eye and that they are tight)
Wash Hamilton syringe 10 times with RO water and 10 times with Acetone.
Spray table and all instruments with 70% EtOH and then with fresh 10% bleach.
Wash and autoclave surgical tools.
Post-op
Put mouse in clean cage with chow on the floor cage and a water bottle. The cage should be on a heating pad in low.
During recovery from anesthesia, observe mice carefully (at least every five minutes).
4-6 hours after Buprenex injection, inject mouse with subcutaneous Rimadyl (5 mg/kg); Stock injection is 0.5mg/mL (for a 30g mice volume injected is 0.3mL)
Once the mice are returned to their holding room, they will be monitored on a daily basis for signs of pain or distress: including decreased activity, hunched posture, decreased grooming, and teeth chattering. Signs of infection, like heat, swelling, redness, and exudation will also be monitored for.