Sep 12, 2022
Stereotaxic rat brain surgery for Substantia Nigra targeting
- 1Vall d'Hebron Research Institute
Protocol Citation: Marta Gonzalez-Sepulveda, Joan Compte, Miquel Vila 2022. Stereotaxic rat brain surgery for Substantia Nigra targeting. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gpj8zpgzp/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: September 12, 2022
Last Modified: November 10, 2023
Protocol Integer ID: 69885
Abstract
Stereotaxic rat brain surgery
Anesthetize rat with isofluorane (5% induction phase, 2% maintenance phase)
Weight rat & shave the head
Inject meloxicam (0.5 mg/mL) and Baytril (5 mg/mL)
Place rat head gently with ears bar onto the stereotaxic equipment
Scalpel + Scalpel holder = incision middle of rat head
Turn vacuum on
Place 4 hemostats in each extremity of the incision (opening in square shape)
Add hydrogen peroxide. Clean. Add saline . Clean.
Remove remaining blood with cotton swab
Remove tissue of the skull with the not-cutting side of scalpel (scratch in order to see bregma and lambda as better as possible, can bleed for a prolonged time, apply saline as necessary)
Check skull horizontality: place drill (adjust with the yellow tool) + take DV in lambda and bregma by placing the drill on the cranium (neither above nor pressing it). If not leveled (+/- 0.1 between Bregma and Lambda), adjust as necessary
Drill very slowly. Regularly check the depth of craniotomy.
Inject vector solutions using a 10 µL Hamilton syringe fitted with a glass capillary
Go to Bregma with the Hamilton syringe
Calculate SN coordinates: AP (5.2), ML (2.0), DV (7.6)
Perform infusion at 0.4 µL/min and let the needle in place for 4 minutes
Seal craniotomoy with bonewax using a small spatula
Sew the incision 3-5x
Add skin glue to incision site