Stereotactic Injections in Mouse and Rat

Remove hair and put 2% on top of the head and into the ears.

Put Vidisic on the eyes.

Put a chip under the skin to mark the animal.

Fix the animal into the stereotactic apparatus (use the mouse adaptor for mouse and only the ear clamps for rat). Put a tissue over the animal to keep it warm during surgery.

Make sure that the left and right side of the skull is positioned as straight as possible (ears).

Use 1% to clean the top of the head and make an incision with a scalpel.

Clean the skull with a spatula and saline and let dry until bregma and lambda are clearly visible.

Check volume and injection speed of the pump, rinse Hamilton syringe with RBS, ETOH and PBS. (Coat the syringe by taking a full syringe of vector and discard in eppendorf). Put vector in the syringe (± 1.5 µl more than you want to inject) and make sure there are no air bubbles.

Make sure bregma and lambda have the same height, correct the position of the head if there’s a difference of more than 0.02cm.

Put the needle at the right position using bregma as a reference (find the right coordinates using the stereotactic atlas of mouse or rat).

Drill a small hole into the skull at this position until the dura mater is visible.

Make a small hole into the dura mater using a thin needle.

Put the Hamilton needle at the right position (go down slowly to prevent tissue damage) and wait for 0h 1m 0s.

Inject vector at max. 0.25 µl/min.

Wait 0h 5m 0s after injection in order to let the vector diffuse into the brain.

Remove the needle slowly.

Close the skin and disinfect with joodalcohol.

Rinse Hamilton syringe with RBS, ETOH, PBS and AD.