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Handling of Mice
Lift the mouse by grasping the tail at the caudal end with the left hand, and allow
it to grip the wire mesh of the cage with its forelegs. Grasp the nape of neck
between the thumb and the index finger. Place the tail from the hand to the left
small finger so that the mouse is held tightly in the left hand. The mouse is now
ready for injection.
A. Oral Admiinisration
Administration by a stomach tube or by an oral feeding needle. The mouse is
held firmly and the feeding needle is held well towards the back of the mouth and
near the upper palate. The needle is than passed-gently-not-forced down the
esophagus. As it passes into the stomach (about 2 inches down) the animal will
exhibit a definite and characteristic gagging. As much as 0.5m1/10 gm of animal
may be administered.
B. Intravenous Injection
The mouse is to be held firmly by wrapping it in a cloth or by placing it in a
suitable animal holder. With a little practice the tail veins can be recognized
readily. It is helpful to dilate the veins by warming before attempting the
injection. Employ a sharp, 26-gauge needle and slowly inject 0.9% saline,(10
ml/kg of body weight) into the mouse. If the needle is not in the vein, resistance
will be felt while injecting and blanching will be observed in the surrounding
connective tissue.
C. Intraperitoneal Injection
This is a common method to introduce drugs into animals. The drug is injected
intothe peritoneal cavity where absorption downward, inject into the lower half of
the abdomen; intraintestinal or intravisceral injection may be avoided. Inject
0.9% saline, (10 ml/kg of body weight) into the mouse.
D. Subcutaneous Injection
The drug is to be introduced directly underneath the skin. A 3/4 -l inch 26-gauge
needle is employed. Insert the needle to its full length to avoid loss of liquid upon
withdrawal. Inject sample of interest (0.1 ml/l0 gm of body weight) under the
skin of a male mouse followed by gentle massage of the site of injection. The
common site employed is the dorsal neck area and that of the abdomen.
E. Intramuscular Injection
Inject sample of interest into the gluteus maximus muscle of a mouse, using a
26-gauge needle. One must always pull back on the plunger to make certain the
needle is not in a vein.
Introduction
Blood is collected from laboratory animals for various scientific purposes, for
example, to study the effects of a test drug on various constituents, such as
hormones, substrates, or blood cells. In the field of pharmacokinetics and drug
metabolism, blood samples are necessary for analytical determination of the drug
and its metabolites. Blood is also needed for some in vitro assays using blood
cells or defined plasma protein fractions. The techniques for blood collection
depend on specific factors which differ from one experiment to the other. There is
a difference between terminal and nonterminal blood collection techniques. The
conditions of blood collection at the end of an experiment which includes death
of the animal (terminal experiment) are completely different (anesthesia, volume
of blood) from those of single or repeated blood collections from a conscious
animal. Terminal blood collection under anesthesia allows the use of techniques
which are not acceptable for non-terminal blood collections.
This technique normally recovers a few drops of blood, adequate for hemoglobin,
microhematocrit and cell counts. Larger blood samples can be obtained by
making a small incision over the vessels 0.5 to 2 cm from the tail base using a
scalpel blade. One half to one milliliter of blood can be withdrawn using this
method. Anesthesia or sedation should be used.
Number of samples: No more than four blood samples should be taken within
any 24-hour period.
Sample volume:10 ul
Adverse effects :
• Infection <1%
• Haemorrhage
Retro-orbital bleeding
Blood sampling by orbital puncture is a technique routinely applied in most
laboratories. The puncture of the orbital venous plexus is often performed in tail-
less animals, e.g. hamsters. This technique is also used in rats and mice, when
larger volumes are required which cannot be obtained from the tail vein.
Basically, retro-orbital bleeding should always be performed under anesthesia.
Pasteur pipettes, micropipettes or microcapillary tubes are used and pushed with
a rotating movement through the conjunctiva laterally, dorsally or medially of the
eye to the back wall of the orbit. Sterile capillary tubes and pipettes are
recommended for use to help avoid periorbital infection and potential long term
damage to the eye. The edges of the tubes should be checked for smoothness to
also decrease likeliness of eye damage.
Although the procedure may appear to members of the lay community as unduly
distressful, the NIH ARAC has determined that in the hands of a skilled
technician retro-orbital bleeding is a humane procedure that produces minimal
and transient pain/distress. Retro-orbital bleeding can be conducted in awake
mice. Due to pain and distress issues retro-orbital sampling in the rat is best
conducted under general anesthesia Rapid and large number of mice/rats can be
bled within a short period of time.
Cardiac puncture
In tailless animals such as guinea pigs and hamsters, cardiac puncture under
general anesthesia may be the preferred technique.
The collection of blood by cardiac puncture has been performed in guinea pigs,
gerbils and hamsters. In these species it is difficult to collect blood by alternative
methods except retro-orbital bleeding. In general, cardiac puncture should be
performed under general anesthesia with atropine as premedication to prevent
cardiac arrhythmia. If cardiac puncture is used for a non-terminal blood
withdrawal with recovery, the animal has to be separated from other animals
until it is fully conscious. It should be carefully watched for adverse effects and
sacrificed if found in distress due to complications like bleeding into the
pericardium or into the thorax.
Sample volume: Up to 1 ml
Conclusion: The given mice were handled the various routes of administration of
drugs were observed and practiced. Also blood collection was preformed using
the tail snip and retro orbital bleeding bearing in mind the volume constrains for
the methods in question.