4. Guarantee the efficacy of the sedation by testing the mouse
reflex to pain (i.e., by pressing the animal’s feet and obtaining
no reaction).
5. With the help of a forceps, hold the tongue maintaining the
mouth open and keeping the sublingual region clear (Fig. 5).
6. With a micropipette, administer the dose (maximum of 10 μL
per administration) under the tongue.
7. Wait until the dose is completely absorbed before repeating the
administration.
8. To avoid swallowing, keep the mouse laying on its abdomen
until the effects of the anesthesia has passed.
9. Repeat steps 6 and 7 until the complete dose volume has been
administered.
Fig. 3 Two-hand mice restraint. With the dominant hand pick the mouse by the base of the tail (a), with the
nondominant hand press the mouse gently against the surface and, with the thumb and index finger, hold the
skin of the back of its neck (b), raise the mouse and, with the dominant hand, tuck the tail between third and
fourth finger of the nondominant hand (c) and the restraining is completed (d)
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