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Anticoagulants are the chemical agents which prevent the coagulation of blood
iv. Heparin
v. Sodium fluoride
3. What is EDTA and what are its different forms and what is its mechanism of action
i. Tripotassium EDTA
Tripotassium EDTA is more soluble and it is more preferred than the other forms.
Mechanism of action: EDTA acts by Chelation of calcium ions/ removing the free calcium ions.
We know that calcium is very much needed for clotting of blood. Hence, when the calcium ions
i. The morphology of the blood cells are well preserved up to 4 hours and hence it is the
Disadvantages
i. If there is excess EDTA/ the volume of blood filled is less so that the relative volume of EDTA is
excess, then it results in shrinkage of RBC’s. This shrinkage can result in erroneously high MCHC
ii. Platelets can also disintegrate and can result in erroneously high values.
platelet clumping which results due to EDTA dependent agglutinins. These are IgG, IgA and IgM
subtypes. As a result of clumping, the platelet clump is read as single giant platelet or even a
small lymphocyte by the automated analyzers and give the reading of falsely low platelet
counts.
Hence in the laboratory it is important to study the peripheral smears for platelet clumps, if the
Double oxalate
7. What is wintrobe’s salt
This is basically a double oxalate. It is a combination of ammonium and potassium oxalate in the
ratio of 3:2 ( 1.2mg and 0.8 mg respectively in 100 ml of distilled water). 0.5 ml of this solution
The oxalates combine with calcium in the blood to form insoluble precipitate of calcium
oxalate. Thus there is depletion of calcium which is necessary for coagulation of blood.
Ammonium oxalate causes swelling of the cells, whereas potassium oxalate results in shrinking.
So, the combined effect is that the cells retain their original shape and size.
Sodium citrate
The sodium citrate binds with the free calcium resulting in sodium citrate complex. Thus
i. Used for estimation of ESR by Westergren’s method. The ratio of citrate and blood has to be
maintained at 1:4
ii. Used in coagulation studies. The ratio of citrate and blood has to be maintained at 1:9
Heparin
13. What is the mechanism of action of heparin
Heparin an acid mucopolysaccharide, has an affinity for blood proteins and act as antithrombin
anticoagulation
i. The blood smear examination is difficult as heparin results in staining artifact. The background
looks pink.
ii. It cannot be used as anticoagulant for cell counts as heparin causes clumping of cells resulting
in erroneous counts.