How do you collect mouse serum?
To obtain the serum from a mouse blood sample I perform this protocol:
- Obtain the sample withouth any anticoagulant and transfer to a strile empty tube.
- Leave the tube in a standing position and wait 30 min.
- Centrifuge 1500 g 10 min at 4ºC.
- Take out the serum.
- Keep the sample at 4ºC up to 6 months.
How do you collect serum from rat blood?
To collect blood serum, place samples (without anticoagulant) at room temperature for up to 30 min to enable clotting. Spin the collection tubes in a refrigerated centrifuge (4 °C) at 2,000 x g. The serum may then be eluted.
How do you collect whole blood from mice?
So-called “terminal” or “non-survival” blood collection is done either pre- or post-mortem and only after the mouse has received surgical anesthesia. Terminal collection can be taken via cardiac puncture and can collect the maximum amount of blood. It can also be done via the aorta or vena cava.
How can we prevent hemolysis during blood collection of mouse?
To avoid hemolysis, blood samples should be taken immediately after the vein has been raised. Avoid “pumping or milking” the blood from the vein as this can induce coagulation and erythrocyte lysis. Avoid high negative pressure in the syringe, as this can cause erythrocytes to rupture.
How much serum does a mouse have?
Under ideal conditions, a skilled person can obtain approximately 50-75% of a mouse’s total blood volume (3-4% of BW) by exsanguination. This amounts to 0.04-0.06 ml blood/gm BW, or 1.0-1.5 ml blood from a 25 gm mouse.
How do you collect serum and plasma?
The clot is removed by centrifugation and the resulting supernatant, designated serum, is carefully removed using a Pasteur pipette. Plasma is produced when whole blood is collected in tubes that are treated with an anticoagulant. The blood does not clot in the plasma tube. The cells are removed by centrifugation.
How do you isolate plasma and serum?
How to separate serum and plasma from blood. Serum is the liquid fraction of whole blood that is collected after the blood is allowed to clot. The clot is removed by centrifugation and the resulting supernatant, designated serum, is carefully removed using a Pasteur pipette.
How much serum is in 5 mL of blood?
The 5 mL red top tube will yield approximately 2.5 mL serum after clotting and centrifuging. Label the specimen appropriately.
How can you separate serum from blood without a centrifuge?
Plasma or serum can be separated from whole blood without centrifugation by allowing the blood to just let stand. By gravity all the cells will settle down in due course of time (if time is not the question). If you allow the citrated blood to stand in a tube, the supernatant is the plasma.
How do you collect plasma from mice?
Plasma can easily be obtained by placing whole blood into a tube containing an anticoagulant. When using a tube with an anticoagulant, do not shake the contents. Instead gently invert the tube end over end several times, watching the blood move through the tube, to ensure adequate mixing.
When to remove cells from a serum sample?
The cells and clotting factors must be removed from the blood sample by allowing adequate time for a clot to form. Most manufacturers of collections systems for serum samples recommend 30–60 min at room temperature for a clot to form and longer if the subject was taking any kind of anticoagulant at sample collection.
How is the protein concentration of mouse serum determined?
The protein concentration of serum was determined with the Bradford protein assay (Bio-Rad Protein Assay Dye Reagent Concentrate, Bio-Rad), using bovine gamma globulin as the standard. The protein concentration ranged from 80 to 90 μg/μl for wild type mouse serum samples.
Which is the best way to collect blood from a mouse?
Cardiac puncture or blood collection from the posterior vena cava are good methods of terminal blood collection when an aseptic sample is needed. collecting blood from the tail in mice over 28 days old and from the orbital sinus in mice of all ages.
How is retro orbital sampling used in mice and rats?
Retro-orbital sampling can be used in both mice and rats by penetrating the retro-orbital sinus in mice orplexus in rats with a sterile hematocrit capillary tube or Pasteur pipette. Sterile tubes are recommended tohelp avoid periorbital infection and potential long-term damage to the eye.