小鼠血液收集SOP

Guidelines: The procedures described in this SOP should be followed when collecting blood samples from mice, rats, or hamsters. Blood volume of an individual animal is approximately 6-8% of body weight in adults. Up to 10% of the total blood volume can safely be removed from a healthy animal without fluid replacement at two week intervals. If a larger sample is needed at one time, up to 15% can be slowly removed if IV fluids are given to replace the lost volume. Maximum withdrawal volumes must be followed to minimize the risk of anemia and cardiovascular collapse.

Body weights can vary with age, strain, and gender. Therefore, it is recommended that animals are weighed to calculate maximum blood collection volumes. For experimental groups of the same age, strain, and gender the average group weight can be used to calculate blood collection limits.

Materials:

Sterile Hematocrit tubes

Sterile Pasteur pipets

Sterile needles- 25G and 23G

Sterile syringes-1CC, 3CC, 5CC and 10CC

Sterile scalpel blades- size 15

Clippers

70% ethanol

Gauze

Blood collection tubes

Heat lamp

Tissue glue

Topical ophthalmic anesthetic, such as proparacaine

Ophthalmic antibiotic ointment

Rodent restrainer

Rat restraint board for jugular blood collection with attached leg restraint ropes

Plastic nose cone for head restraint

Procedures in the mouse can generally be performed by one person. For rats and hamsters some techniques may require two people, one to restrain the animal and one to collect the blood. Other procedures will require a restraint device or light anesthesia. Terminal blood collection by cardiac puncture can only be performed under anesthesia.

Methods:

Tail nick:

This procedure can be used to collect blood from mice.

1.Place a heat lamp over the cage of animals for no more than 5 minutes to warm mice and

dilate vessels.

2.Once the animals are warmed, place them in an appropriately sized restraint device with the

tail extended. An alternative method to using the heat lamp is to place the animal in a

restraint device, then rub the tail for several seconds with a gauze pad soaked in very warm, but not hot, water to dilate the vessels.

3.Wipe the tail with 70% alcohol and allow to dry.

4.Holding the end of the tail, gently twist the tail to expose the lateral tail vein on either side.

5.Approximately 2-3 cm from the tip of the tail, make a small nick over the lateral tail vein

using a sterile scalpel blade. When the nick is made deep enough, blood should start welling up from the nick immediately.

6.Blood can be collected either by touching a capillary tube to the bead of blood, or by

allowing the blood to drop into a collection tube. The tail can be gently stroked from the base of the tail toward the tail to encourage blood flow.

7.Once a sufficient amount of blood is collected, apply pressure to the nick with clean gauze

for 15-30 seconds to stop the flow of blood.