PACKED CELL VOLUME OF WHOLE BLOOD:

MICROHEMATOCRIT METHOD
 HAEMATOCRIT
(Packed Cell Volume)

 PCV is a measurement of the ratio of the

volume occupied by the RBCs to the volume
of whole blood in a sample of capillary or
venous blood.
 PCV is used in evaluating and classifying the
various types of anaemias according to the
red cell indices; used to detect polycythemia,
haemodilution/haemoconcentration.
 PCV is the percentage of RBCs in a volume of
whole blood; it is expressed as percent or
ratio.
 Hb x 3 = Hct ± 3 units
Spun microhaematocrit tube
 Methods for Measurement

1. The spun microhaematocrit method;

2. The automated haematocrit method.
SAMPLE
 Venous blood anticoagulated with EDTA or freely
flowing capillary blood may be used; plain uncoated
microhematocrit tubes must be used.
 With capillary blood, microhematocrit tubes coated
with heparin are used;
 Blood that has been properly anticoagulated with
EDTA is used for automated analysis.
Equipment for Microhematocrit Capillary Tubes
 Special non-graduated capillary tubes
are used; these tubes are 1 mm in
diameter and 7 cm long. Microhaematocrit Microhaematocrit
tube tube sealant
 Some type of seal is needed for one
end of the tube before it can be
centrifuged.
Centrifuge
 A special microhaematocrit
Placement of
centrifuge is used, capable of Microhaematocrit capillary tubes in
centrifuge microhaematocrit
producing centrifugal fields up to centrifuge
10,000 g.
Reading Device
 A special reading device is used to
measure the percentage of packed
RBCs after centrifugation.
Microhaematocrit reader
Reading a haematocrit tube
PROCEDURE
1. Well-mixed anticoagulant blood should be drawn into two
microhematocrit tubes by capillary action; free-flowing capillary
specimens should be collected directly into heparinized capillary
tubes; the tubes should be filled to about three-fourths of their
length. Wipe off the outside of the tubes with a suitable wipe.
2. Seal one end of each tube with a small amount of claylike material
3. Place the filled and sealed capillary tubes into the centrifuge; record
the position number of each specimen.
4. Securely fasten the flat lid on top of the capillary tubes; close the
centrifuge top and secure the latch.
5. Set the timer for 5 minutes; the fixed speed of centrifugation should
be 10,000 to 15,000 rpm.
6. Within 10 minutes, read the microhaematocrit on a reader; measure
the microhaematocrit by adjusting the top of the clay sealant to the 0
mark and reading the top of the red cell column. Do not include the
buffy coat in reading the packed RBC column.
REPORTING RESULTS
 Haematocrit values are generally expressed as a
percentage; with the SI system, PCV is preferably
expressed as a decimal.
 The units are undesignated, but units of liter per liter
(L/L) are implied, so a 45% result is reported as 0.45.
REFERENCE VALUES
 Reference values for Hct are influenced by age, gender,
and altitude and vary among authors.
 Generally accepted values at sea level are:
Adult male, 41.5% - 50.4%
Adult female, 36% - 45%
 Precautions and Technical Factors

 The blood sample must be properly collected and

preserved; any capillary blood samples collected should
be freely flowing.
 Anticoagulated blood samples using EDTA should be
centrifuged within 6 hrs of collection.
 The blood must not be clotted or haemolyzed for any Hct
test.
 Centrifugation must be sufficient to yield maximum
packing of the red cells.
 The Hct value is frequently accompanied by a Hb
determination; the Hct result in percent units should be
approximately three times the Hb result.
 The capillary tubes must be properly sealed so that no
leakage occurs.
 These tubes are not calibrated, so the level of packed RBCs
and the total volume of the cells and plasma must be
accurately measured by some convenient reading device.
 The buffy coat layer is not included in the reading for the Hct.
 Even with adequate centrifugation, a small amount of plasma
remains trapped around the cells; inadequate centrifugation
will give falsely high results.
 Hct values determined by automation are consistently lower
than that done by the spun microhaematocrit method.
 The microhaematocrit will be falsely low because of cell
shrinkage.
 If the tubes are not sealed properly, falsely low results will be
obtained because more RBCs will be lost than plasma.
THE RED CELL INDICES
 INTRODUCTION
 The RBC indices are the mean cell volume (MCV), mean
cell haemoglobin (MCH), and the mean cell haemoglobin
concentration (MCHC).
 The MCV represents the volume/size of the average
RBC, the MCH represents the weight of Hb in the
average RBC, and the MCHC represents the Hb
conc/colour of the average RBC.
 Red cell distribution width (RDW); a measurement of the
degree of variability in RBC size.
 Automated multiparameter instruments measure the
Hb, MCV, and RBC count, then automatically calculate
the Hct, MCH, and MCHC.
 It is important to verify all indices against observations
of stained blood films.
 The units in which the RBCs are measured and recorded
are micrometers (μm) and picograms (pg).
 Red cell indices are calculated from the following
haematology data in units as indicated:

Test Name Abbreviation Units

Haematocrit Hct %
Packed cell volume PCV L/L
Red cell count RBC ×1012/L
Haemoglobin Hb g/dL
 MEAN CELL VOLUME
 The MCV is the average volume of an RBC in femtoliters
(fL).
 The MCV is calculated manually by dividing the volume of
PCV (Hct) by the number of red cells, using the formula:
MCV (fL) = Hct x 10
RBC
EXAMPLE: If the Hct is 45% and the red cell count is 5x1012/L:
MCV = 45 x 10
5
= 90
 The MCV in normal adults is between 80 and 96 fL.
 The MCV indicates whether the RBCs will appear small
(microcytic), normal (normocytic), or large (macrocytic).
 The chief source of error in the MCV is the
considerable error in the manual red cell count, if
used.
 With automated cell counters and electronically
calculated indices, the MCV is measured directly, and
the haematocrit is calculated from MCV and red cell
count (Hct = MCV x RBC).
 MEAN CELL HAEMOGLOBIN

 The MCH is the content of Hb in the average RBC; it is

measured in picograms.
 The MCH is obtained by dividing the Hb by the RBC count:
MCH (pg) = Hb x 10
RBC

EXAMPLE: If the haemoglobin is 15 g/dL and the RBC is 5x1012/L:

MCH = 15 x 10
5
= 30 pg
 The normal range for the MCH is 27 to 33 pg.
 MCH should always correlate with the MCV and the MCHC.
 The chief source of MCH error is the RBC count, if done
manually.
MEAN CELL HAEMOGLOBIN CONCENTRATION

 The MCHC is the average Hb concentration in a given

volume of packed red cells; it is expressed as grams per
deciliter.
 MCHC may be calculated from the MCV and the MCH or
from the Hb and Hct values by using the following formula:
MCHC (g/dL) = MCH x 100 or Hb x 100
MCV Hct
EXAMPLE: If the Hb conc is 15g/dL and the Hct is 45%:
MCHC = 15 x 100
45
= 33.3g/dL

If a PCV of 0.45 is used in the example above:

MCHC = 15 = 33.3g/dL
0.45
 Reference values range from 33 to 36 g/dL, and
values below 32 g/dL indicate hypochromasia.
 An impossibly high MCHC (>40 g/dL) also could
indicate the presence of cold agglutinins in the
specimen.
 The MCHC typically increases only in spherocytosis.
 RED CELL DISTRIBUTION WIDTH

 The RDW is a measurement of the degree of

anisocytosis present, or the degree of variability in RBC
size, in a blood sample.
 In Coulter instruments, a red cell histogram is plotted,
and the RDW (%) is defined as the coefficient of
variation of the MCV:
RDW (%) = Standard deviation (SD) of MCV x 100
Mean MCV
 The reference range for RDW is from 11% to 15%.
PRECAUTIONS AND TECHNICAL FACTORS

 Any manual RBC count, Hct, or Hb conc used in the

calculations must be accurate.
 Check the appearance of the RBCs in a well-stained
blood film against the calculated indices.
 The MCHC is often below 30 g/dL before
hypochromasia is observed on the blood film.
 CASE STUDY
An 80 year old man is seen for an annual physical
examination. He complains of shortness of breath on
exertion and is often tired. His stool is black.
Laboratory data are:
Haemoglobin: 8.2 g/dL
Haematocrit: 30%
White cell count: 4.2 x 109/L
White cell differential:
Neutrophils: 60%
Lymphocytes: 31%
Monocytes: 7%
Eosinophils: 2%
Basophils: 0%
Red cell count: 4.0 x 1012/L
RDW: 20%
Platelet count: 400 x 109/L
Reticulocyte count: 1.2%
Stool occult blood: Positive
QUESTIONS
1.What is this patient’s MCV? Show calculation
2.With reference to the MCV normal range, comment on
the MCV obtained above.
3.How would you classify this anaemia morphologically?
4.What is the most probable type of anaemia seen in this
patient?
5.What is the likely cause of the complaints of shortness
of breath reported in this patient?
END