Biology Reference
In-Depth Information
5. Activate the isofl urane vaporizer. First adjust to 2 %, then
maintain at a working concentration of 1-1.5 % throughout
the procedure.
6. Monitor breathing and check toe-pinch refl ex:
Monitoring of breathing and toe-pinch refl ex should be
done throughout the surgery. Toe-pinch refl ex should be
checked every 10 min but should be omitted when the
injection needle is in the brain (see below).
●
7. Head-fi x mouse:
●
Position ear bars: Bars should be positioned either inside
the ear canal or slightly ventral and anterior to it, where
jaw meets skull. For ideal positioning, both bars should be
pushed against the skull simultaneously and from the same
starting position (
see
Note 5
; Fig.
1
-bI)
●
Adjust the height of the tooth bar and/or ear bars in order
to achieve correct placement of the skull with respect to
bars (
see
Note 6
).
●
Once correct position has been obtained, tighten ear bars
and make sure the head is fi xed tightly by gently applying
pressure to the head in a downward direction.
8. Heating pad:
Heating pad should be placed under the mouse and set
to 37 °C.
●
Body temperature should be monitored using a rectal probe.
!
●
Make sure heating pad does not overheat.
9. Sterilize scalp:
Use sterile cotton swabs to wash with iodine antiseptic,
then with 70 % ethanol. Repeat this step twice.
●
Stroke swab from center of the skull towards the sides.
●
10. Make incision:
Using fi ne-tipped tweezers pull a bit of skin upwards from
skull between the ears of mouse and make a small hori-
zontal incision with the scissors, making sure not to dam-
age other tissues.
●
Insert scissor tips through the cut so scissors are perpen-
dicular to the skull and facing forward. Cut along the
midline until incision reaches between the eyes.
●
Use the bulldog clamps to clamp the skin on each side of
the incision and move laterally to reveal the entire skull
(Fig.
1
-bII)
●
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