Stereotaxic rat brain surgery for Substantia Nigra targeting

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