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    Stereotaxic rat brain surgery for Substantia Nigra targeting

    Miquel Vila, Joan Compte, Marta Gonzalez-Sepulveda

    Published: 2022-09-13 DOI: 10.17504/protocols.io.dm6gpj8zpgzp/v1
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    Abstract

    Stereotaxic rat brain surgery

    Steps

    1.

    Anesthetize rat with isofluorane (5% induction phase, 2% maintenance phase)

    2.

    Weight rat & shave the head

    3.

    Inject meloxicam (0.5 mg/mL) and Baytril (5 mg/mL)

    4.

    Place rat head gently with ears bar onto the stereotaxic equipment

    5.

    Scalpel + Scalpel holder = incision middle of rat head

    6.

    Turn vacuum on

    7.

    Place 4 hemostats in each extremity of the incision (opening in square shape)

    8.

    Add hydrogen peroxide. Clean. Add saline . Clean.

    9.

    Remove remaining blood with cotton swab

    10.

    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)

    11.

    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

    12.

    Drill very slowly. Regularly check the depth of craniotomy.

    13.

    Inject vector solutions using a 10 µL Hamilton syringe fitted with a glass capillary

    14.

    Go to Bregma with the Hamilton syringe

    15.

    Calculate SN coordinates: AP (5.2), ML (2.0), DV (7.6)

    16.

    Perform infusion at 0.4 µL/min and let the needle in place for 4 minutes

    17.

    Seal craniotomoy with bonewax using a small spatula

    18.

    Sew the incision 3-5x

    19.

    Add skin glue to incision site

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