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Hematology Lecture: SPECIMEN COLLECTION

Anticogaulants
1.
2.
3.
4.

EDTA
Sodium citrate
Heparin
Oxalates

EDTA
Chelating agent (calcium-binding)
Choice for CBC & sizing
Less shrinkage of RBCs
Less increase in cell volume
upon standing (3-4hrs)
Prevent platelet
aggregation
1. K2 EDTA (Sequestrene) &
Na2 EDTA (Versene)
2. K3 EDTA
Concentrations of 1.5 mg/1 mL of
whole blood
Proper ratio
Excessive EDTA: shrinkage
of RBCs (e.g. falsely
decreased Hct & ESRblack top tube)
After 6-24 hrs: increase Hct &
MCV
Blood smears after 3-4 hrs:
swelling of RBC, platelet
satellitosis

Osmosis in RBCs
Isotonic equal water intake and
leakage
Hypotonic water seeps in
Hypertonic water leaks out
Complete Blood Count
Hemoglobin
Hematocrit
Total leukocyte count
Total RBC count
Leukocyte differential count
Platelet count

Sodium Citrate
Coagulation studies
Removes Ca by precipitation
Combining with Ca forming
double salt
Concentration of 3.2% solution
Ratio = 1:9
Concentration of 3.8%
Falsely prolonged clotting time
with PT & APTT in slightly
underfilled & high Hct samples
PT, APTT, ESR (1:4 of 3.8%)
Acid Citrate Dextrose (used in
Blood Bank)
Longer shelf life
Preserve RBCs in vitro for
longer survival time for
blood transfusion &
hemolytic processes
Citrate Phosphate Dextrose (BB)
Adisol preservative added
Trisodium Citrate
Blood coagulation
Platelet studies
Stabilized plasma pH
Heparin
In vitro & in vivo anticoagulant
Inactivates blood clotting factor
thrombin & Factor Xa
Best because of minimal
hemolysis of RBC
Functional studies of WBCs &
immunologic markers
Coat capillary blood collection
tubes
Blood transfusion during open
heart surgery & exchange
transfusions
Blue tipped nonheparinized
capillary tube used for
coagulation studies
Red tipped heparinized
capillary tube
Not satisfactory for blood film
preparation
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Hematology Lecture: SPECIMEN COLLECTION

Morphologic distortion of
platelets & WBCs
Bluish coloration
Never be used for coagulation
0.1 0.2 mg/mL of blood
15 to 20 or 15 to 30 U/mL of
blood
Na Heparin & Li Heparin
OFT
ESR
Hct
Special Chemistry tests

Oxalates
Combines with Ca to form an
insoluble Ca oxalate salt
Potassium Oxalate/ Sodium
Oxalate
Distorts RBC
Shrinks WBC
Not ideal for blood smear,
ESR, Hct
Concentration of 1-2 mg/mL
of blood
Ammonium Potassium
Oxalate
Balanced oxalate/ double
oxalate
Ideal for ESR, Hct, & blood
counts
Not ideal for blood smears
Concentration of 0.5 mg/5
mL of blood
Oxalates are not ideal for blood
smears
1. Nuclear degeneration of
leukocytes
2. Cytoplasmic vacuolation of
granulocytes
3. Crenation of RBCs
4. Pseudolobulation & clumping of
agranulocytes
5. Malformation of cells
6. Artifact formation in the nuclei o
monocytes & lymphocytes

7. Phagocytosis of oxalate crystals

Defibrination
Removes fibrin
Ideal for OFT, acid serum &
hemolysis test
Obtain maximal amount of
serum
Preserves better morphology of
RBC & WBC
For buffy coat preparation
(purple coloration if more
RBCs)
SKIN PUNCTURE
Sites
3rd & 4th finger
Plantar surface of heel & big
toe
Medial & lateral surface
Not deeper than 2 mm
Margin of the earlobe
Baby:
lateral side of heel
medial side of heel
posterior tibial
Applications of Skin Puncture
1. infants less than 6 months
2. in young children
3. in adults with poor veins
extreme obesity
severe burns
geriatric patients
patients with thrombotic
tendencies
Advantages
3rd or 4th finger
1. Easily accessible
2. Easy to manipulate
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Hematology Lecture: SPECIMEN COLLECTION


3. Ideal for peripheral blood
smear
4. Less intimidating
Earlobe
1. Less painful
2. Less tissue juice
contamination
3. More free flow of blood
4. Ideal for searching
abnormal cells
Disadvantages of Skin
Puncture
1. Less amount of blood can
be obtained
2. Blood obtained hemolyses
easily
3. Cannot perform additional
or repeated tests
Sample
Mixture of capillary, venous, &
arterial blood

can contain interstitial &


intracellular fluids
Precautions
Prevention of osteomyelitis
1. Warming the area
2. Select appropriate site
3. Cleansing the skin
4. Puncture should be 2.4 to 3
mm deep (in infants: 1.6mm)
5. Avoid extra pressure
6. Avoid double cuts & previous
sites
7. Reduce number of collections
Sites to avoid in skin puncture
1. Cold and cyanotic areas
2. Inflamed and pallor areas
3. Congested and edematous areas
4. Scarred and heavily calloused
areas

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