Feb 16, 2026
Stereotaxic Viral Injections Into VTA and PnO (No Implant)
- Cristian González-Cabrera1,
- Rukhshona Kayumova2,
- Matthias Prigge3
- 1LIN;
- 2Leibniz Institute for Neuorbiology;
- 3Leibniz Institute for Neurobiology, Magdeburg, Germany
Protocol Citation: Cristian González-Cabrera, Rukhshona Kayumova, Matthias Prigge 2026. Stereotaxic Viral Injections Into VTA and PnO (No Implant). protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp1258gzp/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: February 16, 2026
Last Modified: February 16, 2026
Protocol Integer ID: 243385
Keywords: stereotaxic viral injections into vta, stereotaxic viral injection, stereotaxic delivery of viral vector, stereotaxic delivery, oral pontine reticular nucleus, injection, isoflurane anesthesia, optrode implantation, ventral tegmental area, oral pontine, viral vector, no implant, implantation, pno, reticular nucleus, vta
Abstract
This protocol describes stereotaxic delivery of viral vectors into the ventral tegmental area (VTA) and the oral pontine reticular nucleus (PnO) under isoflurane anesthesia. This version covers injections only (no fiber or optrode implantation).
Materials
- Isoflurane anesthesia system.
- Stereotaxic frame and mouse adaptor.
- Heating pad set to 37 C.
- Micropipettes or nanoliter injector suitable for small-volume injections.
- Surgical instruments and sterile supplies.
- Viral vectors kept on ice during setup.
- Analgesic and supportive care supplies.
Troubleshooting
Step-by-Step Procedure
Induce anesthesia with isoflurane (3-4% induction) and maintain at 1.0-1.5% in room air.
Place the mouse in a stereotaxic frame on a 37 C heating pad. Protect eyes.
Shave and disinfect the scalp. Make a midline incision and expose the skull.
Level the skull at bregma and lambda.
Drill a small craniotomy above the target coordinate(s).
Load viral solution into the injector. Lower to the target DV coordinate slowly.
Inject virus using a slow, controlled infusion.
Wait briefly after injection before retracting the injector to reduce reflux.
Repeat for additional targets as required (VTA and/or PnO).
Close the scalp with sutures or wound clips.
Monitor the animal until fully recovered from anesthesia and provide post-operative care.
Critical Steps
Confirm skull leveling before targeting coordinates.
Use slow injections and a dwell time to reduce backflow.
Keep injection approach consistent across animals and conditions.
Acknowledgements
Step-by-Step Procedure
1. Induce anesthesia with isoflurane (3-4% induction) and maintain at 1.0-1.5% in room air.
2. Place the mouse in a stereotaxic frame on a 37 C heating pad. Protect eyes.
3. Shave and disinfect the scalp. Make a midline incision and expose the skull.
4. Level the skull at bregma and lambda.
5. Drill a small craniotomy above the target coordinate(s).
6. Load viral solution into the injector. Lower to the target DV coordinate slowly.
7. Inject virus using a slow, controlled infusion.
8. Wait briefly after injection before retracting the injector to reduce reflux.
9. Repeat for additional targets as required (VTA and/or PnO).
10. Close the scalp with sutures or wound clips.
11. Monitor the animal until fully recovered from anesthesia and provide post-operative care.
Critical Steps
- Confirm skull leveling before targeting coordinates.
- Use slow injections and a dwell time to reduce backflow.
- Keep injection approach consistent across animals and conditions.