How do you make EDTA for mouse blood collection?
For each animal, take a 1 mL syringe and pull up 1 mL of 0.5 M EDTA. Place a 25g needle on the syringe, remove bubbles, and push the EDTA through the needle back into the original bottle. This will coat the inside of the needle and syringe with EDTA to help prevent clotting during the collection.
How do you make mouse serum?
METHOD
- Isolate the mouse in a small cage or container.
- Place the mouse in a restricted space (Fig.
- Swab a portion of the tail ∼1.5–2 in.
- Incubate the blood for 1 h at 37°C.
- Transfer the tube to 4°C.
- Centrifuge the blood at 10,000g for 10 min at 4°C.
- Recentrifuge the serum a second time for 10 min.
How do you collect serum from rat blood?
Blood Collection Hold the syringe with one hand, and pull the syringe slowly to collect 15 ml of blood from a male rat. Remove the needle from the syringe, and pour the blood into two ice-cold 10-ml sterile test tubes (Figure 1A). Keep the tubes on ice until centrifugation to delay the formation of the blood clot.
How much plasma do you get from a mouse?
Without fluid replacement, approximately 0.007-0.008 ml of blood/gm BW can be safely withdrawn (about 0.18-0.2 ml of blood from a 25-gm mouse); with fluid replacement, approximately 0.014-0.016 ml blood/gm BW can be withdrawn (about 0.35-0.4 ml blood from a 25-gm mouse).
How much quantity of EDTA should be used for 1 ml of blood?
e.g. to 1 ml of blood, you need to add additional 2 ul of EDTA. 10.
What concentration of EDTA is used for collection of blood?
Potassium-containing salts of EDTA are widely used and recommended as blood anticoagulants for cell counting and sizing in haematology laboratories. The optimum EDTA concentration is 4455 mmol per litre of blood.
How do you isolate a mouse serum?
The isolation procedure involves mincing of the caput epididymidis (CPE) of mice and rats to obtain a crude CPE suspension. This suspension is filtered to remove tubular fragments. The filtrate containing sperms and cellular contaminants is trypsinized, followed by sonication.
How do you collect a serum sample?
Collection Procedure
- Collect blood in appropriate serum separator tube.
- Gently invert the tube 8-10 times, and let tube sit in an upright position for at least 15-30 minutes at room temperature to allow the blood to clot.
- Label sample with appropriate study code.
How much blood can you sample from a mouse?
How often can you collect blood from mice?
If a single blood sample is needed, the rule of thumb is up to 10 percent of the total blood volume of the mouse may be safely collected every two to four weeks. Up to 7.5 percent of the total blood volume may be collected every 7 days, and up to 1 percent of the total blood volume may be collected daily.
Why is K2 used for EDTA?
Both dipotassium (K2) andtripotassium (K3) Ethylene Diamine Tetraacetic Acid (EDTA) are widely used anticoagulants. International CouncilStandardization of Hematology (ICSH) and several researchers recommend the use of K2 EDTA due to its less hyperosmolareffect on blood cells compared to K3 EDTA.
What does K3 EDTA stand for?
The tubes are available with interior coated spray-dried K2EDTA (dipotassium ethylenediaminetetraacetic acid) or K3EDTA (tripotassium ethylenediaminetetraacetic acid). Both EDTA salts inhibit the coagulation of the blood. specimen by binding Calcium (Ca2+), thus preserving.
What is the difference between K2 EDTA and K3 EDTA?
The main difference between K2 EDTA and K3 EDTA is that K2 EDTA contains two chelated potassium ions whereas K3 EDTA contains three chelated potassium ions.
What color tube is used for PRP?
Diego how you harvest the platelets depends on what you will do with them afterward. If you need them functional, then drawing into ACD (yellow-top) tubes rather than citrate (blue top) will allow you to pellet the PRP at 2000g without activating the platelets or clumping them.
How do you warm up a mouse before blood collection?
Warming the mouse immediately prior to blood collection will increase blood flow considerably. Place a lamp over the cage for five minutes or place the cage on a heating pad, on the lowest setting. Place the mouse in a restraining tube so its head is covered and its hind legs are free. Grasp the fold of skin between the tail and thigh (Fig. 1).
Why do we collect blood and plasma for mouse phenotyping?
There are many elements in plasma that can act as surrogate markers of the physiological well-being of a mouse, thus making the collection of blood and plasma a general technique with many applications in mouse phenotyping. For example, the presence of certain enzymes in plasma can serve as markers …
How do you take a blood sample from a mouse?
Care must be taken to make sure that the blood does not come in contact with the mouse’s skin or fur. Cardiac puncture or blood collection from the posterior vena cava are good methods of terminal blood collection when an aseptic sample is needed. Volume Required
What is the best method of terminal blood collection in mice?
Cardiac puncture or blood collection from the posterior vena cava are good methods of terminal blood collection when an aseptic sample is needed. Volume Required On average, the total blood volume in the mouse is 6-8% of its body weight, or 6-8 ml of blood per 100 g of body 48 TECHNIQUEVolume 29, No. 10 Lab Animal November 2000