Practical Immunology And Serology Injection in Lab Animals

1. Practical Immunology
And Serology
Injection in Lab Animals
By
Ahmed Riyadh Abdul Rahman
Al-Noor University College
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2. Injection in Lab Animals
 Injection in lab animals are using for many purpose like immunization animal against
any disease, antibodies produced can be used as vaccine for disease.
 Immunize lab animal (ex. mouse or rabbit) by Injection with a specific antigen to
stimulate its immune system.
The aim of immunization
To prepare specific polyclonal immune sera.
For the production of monoclonal antibodies.
Immunization animal against any disease (Protective).
Research into the protective effects of vaccines and vaccine quality control.
Basic immunological research.
Induction of disease models.2

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Laboratory Animals Used for Immunization
Characteristic of Lab Animals Using in Injection
1- Empty of infection (healthy).
2- Specific Weight, age and type.
3- Genetically pure.

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Methods of Vaccine Administration (Injection) Routes in Lab Animals
1-Intravenous (IV) route: injected directly in the vascular
system through a vein.
Procedure
• Use the lateral tail vein located on either side of the tail.
• Warm the mouse under a heat lamp to promote peripheral vasodilation
because the tail vein is difficult to find, being sure not to OVERHEAT.
• Place the animal in a restraint device and stabilize the tail between the thumb
and forefinger of the hand that will not be manipulating the syringe.
• Prep the tail with 70% ethanol.
• Attempt the injection starting at the middle or slightly distal part of the tail.
• Insert the needle, bevel up, approximately parallel to the vein and insert the
needle at least 3 mm into the vein.
• Must inject slowly.

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2-Intraperitoneal (IP) route: injected into the abdominal cavity.
Procedure
 The injection site is usually on the animal’s lower abdominal quadrant.
 Clean injection site with ethanol.
 Insert the needle at approximately 30-45 degree angle.
 The position of injection is in the abdomen, not too high, not too low, if too High, liver may be hurt, if too
low, bladder may be hurt .
 Before injection, Aspirate: if * greenish-brown aspirate indicates needle penetration into the intestines *
yellow aspirate indicates needle penetration into the bladder. If nothing is aspirated, Inject vaccine and
pressure on the area.

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3-Intradermal (ID) route: the vaccine is injected into the top layers of the skin
(between the layers of the skin).
Procedure
 The mouse should be anesthetized, Remove the hair at the injection site.
 Swab the site with 70% ethanol.
 Stretch the skin tight between the thumb and index finger.
 Insert the needle into the skin, bevel up, holding the needle nearly parallel to the plane of
the skin between the epidermis and the dermis.
 Do not aspirate and inject the material slowly.
 Injection of the compound will create a bleb, or small blister, in the skin.
 If the needle is inserted into the subcutaneous space, no bleb is formed. Injecting too
deeply results in a subcutaneous injection.
 Pause after the injection to allow the skin to stretch and adjust before withdrawing the
needle.

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Intradermal (ID) route

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4-Subcutaneous (SC) route: the vaccine is injected into
the area below the skin into the fatty, connective tissue and
above the muscle (under the skin).
Procedure
Most common injection site is the loose skin around the neck and
shoulder area.
• Restrain animal.
• Clean injection site with alcohol.
• Lift the skin over the back to form a tent.
• Insert needle at base of skin fold between thumb and finger.
• Pull back the syringe plunger to aspirate the syringe. Any blood
indicates improper needle placement, and needle must be
repositioned.
• Administer substance in a steady, fluid motion.

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5-Intramuscular (IM) route: the vaccine is injected into the muscle.
Procedure
IM injections are administered in the thigh muscles of the hind limb.
 Restrain mouse by either holding the mouse against your body (and isolating the rear leg or
have a second person restrain while you inject.
 Clean the injection site with alcohol.
 Insert needle into thigh muscles, direct the needle toward tail, it is very important to avoid
injuring the sciatic nerve.
 Pull back the syringe plunger to aspirate the syringe. Any blood indicates improper needle
placement, and needle must be repositioned.
 Slowly inject the material.

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6-Inhalation route: Inspiration through nose or mouth.
7-Intranasal route: administered into the nose.
8-Oral route: the vaccine is given by drops into the mouth.

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9- Pads injection: can be applied in the internal sites of foot.
Note: Not all techniques are appropriate for each species. For example; IM injections
are avoided in rodents because the amount of material that can be injected into the
rodent's limited muscle mass is so small that the technique is not practical; IP injections
are not administered to rabbits as other techniques are more suitable.

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Any question?