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Packed Cell Volume (PCV)

Packed cell volume (PCV), also known as hematocrit, measures the ratio of red blood cells to total blood volume and is used to screen for anemia and other conditions. The microhematocrit method is preferred for determining PCV as it allows for higher centrifugation speeds, shorter times, and less trapped plasma between cells. Proper technique and quality control measures like running controls are important to obtain accurate and reliable PCV values.

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Eyasu demsew
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915 views24 pages

Packed Cell Volume (PCV)

Packed cell volume (PCV), also known as hematocrit, measures the ratio of red blood cells to total blood volume and is used to screen for anemia and other conditions. The microhematocrit method is preferred for determining PCV as it allows for higher centrifugation speeds, shorter times, and less trapped plasma between cells. Proper technique and quality control measures like running controls are important to obtain accurate and reliable PCV values.

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Eyasu demsew
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CHAPTER 10

Packed cell volume (PCV)


Objectives
At the end of this chapter students will be able to:
 Define packed cell volume
 Identify the methods used in hematocrit determination
 List the materials required in PCV determination using
the micro- and macro- hematocrit methods
 Discuss the advantages of the microhematocrit method
of PCV determination
Objectives cont’d
 Discuss the clinical significance of PCV determination
 Indicate the normal hematocrit values in health
 Perform PCV determination on a sample of blood using
the microhematocrit and Wintrobe method
 List the sources of error in PCV determination
 Apply QC measures for PCV determination
Outline
10.1. packed cell volume (PCV)

i. Definition
 Commonly referred to as hematocrit
 is a measure of the ratio of the volume occupied by the
red cells to the volume of whole blood in a sample of
capillary or venous blood
 The ratio is measured after appropriate centrifugation in
the manual methods
 Expressed as a decimal fraction (in l/l) or as a
percentage (%) to the nearest 0.5%
PCV cont’d

 Buffy coat is
composed of
WBC and
platelets
Hematocrit Determination cont’d
Significance of the test
 enables the calculation of the red cell indices that are
widely used in the classification of anemias. These are:
 Mean cell volume (MCV),
 Mean cell Hb concentration (MCHC) and
 to screen for anemia when it is not possible to measure
hemoglobin, and
 to diagnose polycythemia vera and to monitor its
treatment.
Hematocrit Determination
Methods
 Macro method
 Wintrobe
 Micro methods
 Adams microhematocrit method

 Electronic method
 based on the principle that the average red cell volume is
determined, the red cell count made , and the hematocrit
found by calculation e.g. coulter counter
10.2. Microhematocrit Method
iii. Principle of test
 Anticoagulated blood in a glass capillary of specified length, bore
size, and wall-thickness is centrifuged in a microhematocrit
centrifuge at 10000-15000 RPM for 5 minutes to obtain complete
packing of the red cells
 The PCV value is read from the scale of a microhematocrit
reader or calculated by dividing the height of the red cell column
by the height of the total column of blood.

 A small amount of plasma remains trapped between the packed


red cells.
Microhematocrit Method cont’d

 Note: Due to trapped plasma, PCV values using a


centrifugation technique are 1-3% higher than those
obtained from an electronic cell analyzer which
computes the value from the MCV and red cell count
(PCV = MCV x RBC).

iv. Specimen:
 either well mixed EDTA anticoagulated blood or
 capillary blood collected into a heparinized capillary tube
Microhematocrit Method cont’d
v. Equipment
 Microhematocrit centrifuge
 fixed speed (10000-15000 rpm)
microhematocrit centrifuge with
essential safety features which
include
 a lid interlock,

 metallic casing and

 counterbalanced lid and fitted

with a digital timer is required


 HCT reader
Microhematocrit Method cont’d
 Capillary tubes
 plain (blue band) or heparinized (red band) capillaries
 measuring 75 mm in length
 internal diameter of 1 mm and
 wall thickness of 0.2 – 0.25 mm

 Sealant
 plasticsealant, modeling clay, or plasticine.
Note: heat sealing should be avoided since it distorts the end
of the tube resulting in breakage, or the heat damages the red
cells resulting in an incorrect PCV. Do NOT also use soap or
any other sealing substances for sealing Hct tubes since
detergents lyse RBCs
vii. Method
 If capillary blood from finger puncture is to be used, follow SOP for
capillary blood collection
 If anticoagulated venous blood, mix specimen well
 Allow the blood to enter the tube by capillarity
 Fill 3/4th of the capillary tube ; do in duplicate
 Seal the capillary tubes by vertically placing the dry end into a tray of
sealing compound (wax or plasticin)
 Rotate the capillary tube slightly and remove it from the tray. The
sealant plug should be 4-6mm long. Inspect the seal for a flat bottom.
 Place the filled, sealed capillary tube in the groove (slots) of the
centrifuge with the sealed end toward the periphery.
Microhematocrit determination cont’d
 Set the timer of the centrifuge at 5 minute and spin at
10,000-15,000g.
 Read the PCV using a reading device that is either part of
the centrifuge or separate from it

 Alternatively, the ratio of the red cell column to whole


column (i.e., plasma and red cells) can be calculated from
measurements obtained by placing the tube against
arithmetic graph paper
 If no reader at all, use a ruler
Example: height of red cell column = 19mm
height of total blood column = 49mm
PCV = 19mm/49mm = 0.388 (l/l) or 38.8%
vii. Quality control
 For precision check
 Perform in duplicate
 duplicates must agree within 1.5 %

 For accuracy check


 Run a whole blood control with known results

 Note: Only for QC purpose check “Rule of 3”


 Hb x 3= Hct  3%
viii. Sources of Error in Hct Procedures

 Specimen collection errors


 Inadequate mixing of specimen (for EDTA blood)
 Improper use of the hematocrit reader or including the
buffy coat layer
 Improper centrifugation
 Wrong RPMs (speed) of the centrifuge
 Incorrect centrifuge time
 Manually stopping the centrifuge
 wrong sealing method used (e.g use of soap, heating)
 Incorrect reading due to uneven clay plug
 Excess anticoagulant
Sources of Error cont’d
 Incomplete packing due to insufficient centrifugation.
Centrifuges should be regularly checked for proper
operation (use tachometer to check rpm & document)
 Incorrect reading of results.
 Hemolysis or clotting of samples:
 Factors that cause hemolysis and clotting of samples
should be controlled.
 Occasionally, the red cell – plasma interface is not clear-cut
and the hematocrit is difficult to read. In such cases repeat
the test ensuring proper filling and centrifugation.
 Variation of the bore of the tubes cause serious errors if
they are not manufactured within the narrow limits of
precision that conform to defined standards
ix. Interpretation

 Reference range varies between age, gender and altitude


Children at birth 44-54%
Children 2-5 years 34-40%
Children 6-12 years 35-45%
Adult men 40-54%
Adult women 36-46

Ethiopia: Adult Males 41.6 – 55.1%


Adult Females 35.3 – 48.8%
Interpretation cont’d

 Decreased in anemia
 Increased in polycythemia

Limitation
 Plasma trapping results in higher values (1-3% higher
than those obtained from an electronic cell analyzer)
Additional Note
 The technician should cultivate the habit of inspecting
both the buffy coat and the supernatant plasma when
reading the hematocrit value.
 A note should be made on the patient’s report if an
abnormal plasma or buffy coat is seen as this is often an
important clue for the clinician.
 Example:
 very thick buffy coat: may mean cases of leukemia

 very thin buffy coat: may show marked leucopenia

 yellowish plasma: may show jaundice (if pre-analytic

error can be ruled out)


 When a thick buffy coat is seen, perform total and
differential WBC count
10.3. Macrohematocrit method
 Procedure is no longer in routine use
 Wintrobe tube is filled with well mixed EDTA
anticoagulated venous blood to the mark "0" on top using
a long stem Pasteur pipette making sure that no air
bubbles are trapped.
 The ratio of EDTA to volume of blood should be
1.5mg/ml or 0.1ml 10%w/v K3EDTA/ml.
 EDTA inexcess of this proportion may cause a falsely
low PCV as a consequence of cell shrinkage.
Macrohematocrit method cont’d
 The preparation is then spun at 2300g for 30 minutes.
 The hematocrit is read from the scale on the right hand
side of the tube taking the top of the black band of
reduced erythrocytes immediately beneath the reddish
gray leukocyte layer.
Advantages of the Microhematocrit
Method
 It enables higher centrifugation speeds with
consequent shorter centrifugation times and
superior packing.
 The amount of trapped plasma is less than that
in the Wintrobe method by virtue of the higher
centrifugation speed employed.
Review Questions
1. Define PCV. What is the significance of measuring PCV in
a sample of blood?
2. List the possible sources of error in PCV determination
using the microhematocrit method.

3. List the items that are required in PCV determination using


the micro- and macrohematocrit methods.

4. What is the advantage of the microhematocrit method of


PCV determination?
5. How do you relate measured PCV and Hb values of a
sample of blood?
6. What quality control measures do you apply to ensure the
reliability of PCV values?

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