Biomedical Engineering Reference
In-Depth Information
3. Load the syringe with the virus to be injected. We recommend
injecting 2
L into each of the lateral ventricles
a
.
4. Place a neonate pup (0-1 day old) into the conical tube and
lightly tighten the cap. Do not screw it on fully. Watch the pup
for movement. Remove immediately once the pup has stopped
moving.
5. Gently, but securely, hold the pup between your thumb and
index fi nger, and identify the lambda suture landmark. Insert
the syringe tip 0.5 cm ventral, 1 cm lateral from the sagittal
suture, and 2 cm anterior from the transverse suture. Slowly
inject 2
ʼ
L.
6. Repeat for the contralateral ventricle
b
.
7. Place the pup on a warm surface, and wait for it to wake up
from anesthesia
c
.
8. Rub the pup with bedding from its home cage, and place it
back into its cage.
9. Once all the pups have been injected and returned to their
home cage, return the mother as well
d
.
10. Clean the syringe with 0.5 M sodium hydroxide, as noted
above (step 3)
e
.
ʼ
(a) The syringe should be cleaned every time a new virus is used.
Cleaning should be done by loading the syringe with 0.5 M
sterile sodium hydroxide for ten minutes, followed by 15-20
fl ushes with sterile water.
(b) If the sutures are hard to distinguish, place the animal on a dis-
secting illuminator, and illuminate the head from below.
However, be certain that the illuminating surface is not too
hot, as to not burn the animal. Preferably use a fi ber-optic light
to prevent this potential issue.
(c) The pup should wake up within 5 min.
(d) If problems arise with the mother's rejection of the pups, rub
down the animal fi rst with a paper towel moistened with etha-
nol. Likewise, pat the nose of the mother with the same paper
towel. If the pups have been marked in any sort of way (i.e., a
sharpie, tattoo paste), also mark the mother (on the tail or fur).
Note that it is common for the mother to pick up the pups and
carry them to a different spot in the cage.
(e) It is common for the needle to clog, so having a stock of nee-
dles is recommended.
2.2.3 Notes
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