Blood collection

Blood collection
 Blood must be collected with care and adequate safety
precautions:
 Reliable test results
 contamination of the sample is avoided
 infection from blood transmissible pathogens is prevented

N.B All material of human origin should be regarded as capable of

transmitting infection
 wear disposable rubber gloves
 cover any cuts, abrasions or skin breaks on the hand with
adhesive tape and wear gloves
Collection….
 If needle stick injury encountered
 Remove glove
 Squeeze the wound
 Flash with running water
 Scrub the wound with 0.1% NaOCl
 Three general procedures
 Skin puncture
 Venipuncture
 Arterial puncture
Capillary blood collection
 Capillary blood (peripheral blood / microblood samples)
 when only small quantities of blood are required
e.g. Hgb quantitation, WBC and RBC counts
 blood smear preparation
 when venipuncture is impractical
e.g. infants, sever burns, extreme obesity
 Intravenous medication
Capillary….
 Sites of Puncture
 Adults and children
 palmar surface of the tip of the ring or middle finger
 free margin of the ear lobe.
 Infants:
 plantar surface of the big toe or the heel.
Capillary..
 Advantages of Capillary Blood
 It is obtained with ease
 for peripheral blood film preparation(no anticoagulant is
added that
 affect cell morphology)
 Disadvantages of Capillary Blood
 Only small amounts of blood can be obtained
 Platelet count can not be performed
 Precision is poorer (variation in blood flow and dilution
with interstitial fluid)
 Blood in microtubes frequently hemolyses
Venous Blood Collection
 for tests that require anticoagulation or larger quantities of
blood, plasma or serum
 Sites of Puncture
 The veins in the antecubital fossa of the arm are the
 cephalic, median cephalic & median basilic
 In infants and children
 external jugular vein in the neck region
 the femoral vein in the inguinal area
Venous….
 Advantages of Venous Blood
 provide sufficient amount of blood/repeated tests
 Aliquots of the specimen (plasma and serum) may be
frozen for future reference.
 It reduces the possibility of error resulting from dilution
with interstitial fluid
Venous….

 Disadvantages of Venous Blood

 lengthy procedure that requires more preparation
 technically difficult in children, obese individuals and in
patients in shock.
 Needs anticoagulants
 Hematoma (or blood clot formation inside or outside the
veins)
Difference between peripheral & venous Blood
High in peripheral High in venous
 platelet count higher by
 Slightly greater: PCV, RBC
9% in venous than
count and Hgb peripheral blood
 8%: total WBC &
neutrophil counts
 12%: monocyte count by
Arterial puncture
 Arterial blood is used to measure oxygen and carbon dioxide
tension, and to measure pH
 These blood gas measurements are critical in assessment of
oxygenation problems
 pneumonia, pneumonitis, pulmonary embolism
 Arterial punctures are
 technically more difficult
Anticoagulants
anticoagulants
a. Ethylenediamine tetraacetic acid (EDTA)
 the standard hematology anticoagulant
 very efficient and complete anticoagulation
 Lack of effect on the size (morphology) or number of blood
cells
 preferred anticoagulant for cell counts and morphological
studies
 especially for platelet counts and platelet function tests
/prevents platelet aggregation
 Action: chelating ionic calcium

 1.5±0.25mg of Na2 or K3 EDTA/1ml of blood (e.g. 0.02ml of

10% (W/V) solution of K3EDTA is used for 1ml of blood).
Anticoagulants
b. Trisodium Citrate
 combines with calcium, thereby
 Prothrombin X
thrombin
 1:9 and 1:4 for ESR
c. Balanced or double oxalate
 bind and precipitate calcium as calcium oxalate
 3 parts of ammonium oxalate is balanced with 2 parts of
potassium oxalate (neither salt is suitable by itself)
 1-2mg/ml of blood
 Anticoagulants…
d. Heparin
 an excellent natural anticoagulant extracted from mammalian
liver or pancreas
 Action: inactivating thrombin,

 fibrinogen X fibrin
 best when absolute minimal hemolysis is required (e.g., OFT
and HC determination)
 unsatisfactory for leucocyte and platelet counts /cell clumping
 blood film preparation -diffuse blue background
 0.1-0.2mg of the dry salt for 1ml of blood.
Blood Film Preparation
Preparation of blood film
Desirable qualities of a thin blood film
 sufficient working area.
 Acceptable morphology within working area
 Gradual transition to thickness from the thick to thin areas
terminating in a feather like edge.
 No ridges, holes or waves
 Smooth &b continuous margins
 The minimum length of the film should be 3.0cm
(approximately 3/4th of the length of the slide.