AAV Gene Therapy Strategies for Lysosomal Storage Disorders with Central Nervous System Involvement - Gene Delivery and Therapy for Neurological Disorders - page 272

Fig. 3 Stereotaxic frame and injector setup for intracranial injections in adult
mice

1. Set up and connect stereotaxic instrument with injector and
syringe pump controller according to manufacturer's instruc-
tions ( see Fig. 3 ).
2. Attach autoclaved needle to autoclaved syringe a .
3. Prepare a sterile and fresh mix of anesthesia with ketamine
(10 mg/mL fi nal concentration) and xylazine (1 mg/mL fi nal
concentration) using sterile 0.9 % saline to dilute.
4. Anesthetize mouse by intraperitoneal injection of ketamine/
xylazine mix (100 mg/kg mouse dose and 10 mg/kg mouse
dose) b .
5. Apply sterile ophthalmic ointment to the eyes to prevent dry-
ing of corneas.
6. Shave the top of the head using a small electric shaver.
7. Clean the shaved area with a 70 % isopropyl alcohol prep pad
followed by a povidone-iodine prep pad. Repeat three times.
8. Place mouse onto stereotaxic instrument c .
9. Make a longitudinal incision big enough to expose the bregma
(Fig. 4 ) and desired target region d .
10. Open the incision by pushing the skin aside using sterile cotton
swabs, and clean off the periosteum from the skull. Allow skull
to dry e .
11. Load needle and syringe with sterile AAV vector preparation
by hand f .
12. Place needle into injector and stereotaxic instrument accord-
ing to manufacturer's instructions.
13. Align the needle with the bregma on the mouse skull by mov-
ing the stereotaxic manipulator arms in the X, Y axes ( see Fig. 4
for location of bregma) g .

2.1.2 Protocol