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Ch 8.Immunohistoch / immunology
Ch 10.GC/MS, NMR and Proteomics
Tail Vein Injection Techniques (1) (2)
Protocol 1. INTRAVENOUS TAIL VEIN INJECTION PROCEDURE FOR RATS AND MICE
1.
Technician must be in PPE including gown, booties, mask, cap and
gloves.
2. Weigh the rodent prior to
the injection. The injection volume will
be determined by the weight of the animal. The rule of thumb is that the
injection volume
should be no more then 10% of the body weight.
3. The rodents are initially put under a heat lamp
to increase blood
flow to the tail vein. They can be placed in a plastic cage, chamber, or
bowl with the lamp above them. More
then one animal can be heated at a
time.
4. The mice are transferred to a holding device, (AIMS restrainer)
which restrains the mouse while allowing access to the tail vein. The rat
can be placed in a Decapicone(r) or inverted plastic, liter
restrainer
with hole cut out for tail access.
5. The lateral tail vein is identified on either side
and the saline
mix is injected using the smallest syringe possible with a 27g needle for
mice and a 25 g needle for rats. Start
at the tip of the tail and move
closer to the body if you need to stick the animal more then once.
6.
You should be able to see the needle inside the vein or at least
see the fluid go into the vein. Do not insert the needle all
the way to
the hub. Just far enough to get the bevel inside the vein.
7. The needle should be as flat
and parallel as possible to the tail.
The needle can be bent if needle or the tail held below the bench top to
form a level injection
site.
8. After removing the needle, hold the site with gauze to stop the
bleeding before returning to
cage.
9. They can be returned immediately to their cages.
Protocol 2. Mouse Tail Vein Injection (JK Lab 2006)
This is for Adenoviral Delivery System. If you try to deliver plasmids, all procedure is the same for tail-vein
injection, however the material would be quite different. For adenoviral delivery, we make total injection volume 500~600ul with adenovirus
and PBS)
Introduction
Although the mouse is easier to restrain than for the intraperitoneal injection, the tail vein injection
route requires considerable skill in locating the vein and making sure that the needle is inserted into the vein (and not into the
tissue that surrounds the vein). Also, absolutely no air can be accidentally infected, as this may cause air bubble blockage of blood
circulation(and death of the animal).
Procedure
Set up a restraining device and syringe (1ml luer-lok tip syringe + 27G
½ needle) with your material. à no bubble inside the syringe
Prepare at 50C water bath. No more than 55C, otherwise mouse
will be burn.
Remove mouse from the cage by its tail and place mouse into the barrel of the restrainer. Insert the plunger and
push it far enough to make that mouse cannot move.
Put mouse tail into water bath to make it swollen. After 2~3 min, diameter
of vein is expanding.
Locate one of the two lateral veins of tail.
Hold the tail and restrainer at the same time!! à this
is very important !!
Line up(bevel side up) the needle exactly in line with vein !!
Here is the step that takes a lot of
practice to master !!
Insert the needle into vein (sounds easy, doesn’t it!!). If you are successful, you will actually see the tip
of the needle inside of the vein and the injection will be very easy.
If you hit the right position, you can inject more than
2ml very smoothly, otherwise, even 100ul would be very difficult to be injected.
If you are not in the vein, you will still see
a “clearing” of the color around the vein, but only near the injection site. Also, the tissue near the injection site will swell up
and the plunger will not move easily. In that case, stop injecting and “try again” at a site more nearer to the body.
If
you are unsuccessful even after trying different sites on the vein or using the other vein, you have no choice but to return the mouse
to its cage and try a different mouse.
It happens quite frequently that mouse will lose its tail after tail vein injection
failure because of the infections. The tail color will turn to black, and start to be rotten and destroyed. Therefore, you have
to make a note on the cage after tail vein injection, otherwise, Animal facility will report to DAR.
Discard the needle and
syringe. Tail vein injection needs a lot of practice and skill.