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Practical Hematology Manual #1

Practical Hematology Manual #1

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Published by: walhali on Feb 10, 2013
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03/14/2014

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 Al-Azhar University- GazaFaculty of Applied Medical Sciences Laboratory Medicine Department 
Practical Hematology Manual #1
 Prepared by: Ashraf Shaqalaih BSc(MT), MSc(MT), CLS(H), CLSp(H)
Clinical Laboratory Specialist in HematologyClinical Immunohematologist Technologist (Lic#238, State of California, USA)
1
 
 Anticoagulants Used In The Hematology Laboratory 
 
 Anticoagulants
 
are defined as substances which preventblood clotting / coagulation, and allow separation of the blood intocellular and liquid (plasma) components. Generally plasmacontains coagulation factors. The three anticoagulants commonlyused in hematology laboratory are:
1] 
 
thylene
D
i-Amine
etra-Acetic 
 A
cid ( 
EDTA
): 
EDTA can be found in three salt forms:1-Tri-Potassium EDTA2-Di-Sodium EDTA3-Di-Lithium EDTAAlso, EDTA can be crystalline or liquid. Liquid EDTA tubesrequires specific filling volume to avoid dilution effect. So,blood : anticoagulant ratio must be maintained (this is applicableto all anticoagulants). EDTA is also known as
Versene
or 
Sequestrene
. EDTA acts by chelating / removing ionized calcium(calcium is required for blood to clot, so when it is removed bloodwill not clot).
Generally tri-Potassium EDTA is better than di- Sodium EDTA and di-Lithium EDTA.
 
 
 Always, be sure to mix blood with anticoagulant in a manner that guaranteeproper complete mixing, by gentle repeated inversion of the tube, in figure of 8inversion for at least 20 times, do not shake or use vigorous inversion, since thismay cause hemolysis, and disintegration of cells, and the final effect will beerroneous low results for cellular components of blood, which are our hematology laboratory interest.
2
 ANTICOAGULANTS Used in Hematology Laboratory 
 
EDTA is the most commonly used anticoagulant in thehematology laboratory, and is the anticoagulant of choice for theCBC.Excess EDTA (i.e. more EDTA, you fill less blood volume,so EDTA is in excess), causes shrinkage of RBC’s, causingfalsely / erroneously reduced hematocrit (HCT), and subsequentincrease in MCHC and decrease in MCV (MCV and MCHC areRBC indices that will be studied later). Platelets are also affected,they will swell and subsequently disintegrate, causingerroneously high platelet count, since platelets will be disintegratedinto more than one fragment, each fragment will be counted asone platelet (for example if one platelet will be disintegrated into 4fragments, the 4 fragments will be counted as 4 platelets, butactually they represent one platelet, causing erroneously highplatelet count).From the previous discussion we conclude that correct ratio of blood to anticoagulant is very important, to rule out these in vitroeffects.EDTA can induce platelet aggregation and clumping, causingfalsely decreased platelet count, because these platelet clumps willnot be counted as platelets, they may counted as red blood cells(causing low platelet count and high red blood cells counts). Thistechnical problem can be solved by (1) repeated measurements,(2) extraction of new sample and repeat measurements, (3)study the automated cell histograms, and (4) by visualizing bloodfilm, looking for these platelet clumps. Also, Aggregated andclumped platelets interferes with WBC counting zone in
3

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