Sulaymaniyah Polytechnic University

Technical College Of Applied Sciences

Medical Laboratory Science
*First stage – Second Semester
*Lab Technic Report

Supervisor: Prepared By:

 M. Aryan faraj Khushkan Mohammed Mahmood
 M.Syamand Ahmed

( 2019_2020)
Contents:
Introduction:...............................................................................................................................................1
Method of determination..........................................................................................................................2
Procedure:..................................................................................................................................................2
Aim and principle......................................................................................................................................4
Material and reagent.................................................................................................................................4
Normal value and clinical manifestation.................................................................................................5
Discussion...................................................................................................................................................6
References:.................................................................................................................................................7
 Packed Cell Volume
Introduction:
Packed cell volume also known as the hematocrit test, the PCV or Packed Cell Volume Test
is a test done to diagnose polycythemia, dehydration or anemia in certain patients. It is
generally a part of the full blood count test that is used to estimate the need for certain
blood transfusions and monitor the response to treatment. Blood, in general, is a mix of
plasma as well as cells.

The PCV test measures how much of the blood consists of cells. If the PCV returns a
reading of 50%, it means that 50 ml of the cells are present in exactly 100 ml of blood. If the
RBC number increases, then the total reading of the PCV is also up. This number can also
increase due to dehydration.

Performing the PCV tests and the total solids is a pretty routine and simple test undertaken
at many hospitals. All medical members can easily perform the test but interpreting them is
the tricky part. The readings can provide a lot of information regarding the patient's status
and also help plan the next treatment step. The hematocrit test determines the percentage of
whole blood that is made up of red blood cells (RBCs). A normal hematocrit test result for
males is about 45% (38-50%) of blood volume. For females, it is about 40% (35-44%) of blood
volume.

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Method of determination
When anti-coagulated blood is centrifuged at a standard speed, erythrocytes, which are
heavier than white cells and plasma, will settle down at bottom. This red cell volume is
known as Hematocrit or Packed Cell Volume (PCV).

Hematocrit or PCV is the volume of red cells expressed as a percentage of whole blood.

There are two methods, used for the determination of determination of PCV:

 Macro hematocrit method (Wintrobes method): A large volume of blood is required in this
method. Approximately 2 to 4 ml is required.

 Microhematocrit method: This method requires small amount of blood, 2 to 3 drops only. The blood
can be obtained by finger puncture.

Procedure:

 Macro hematocrit procedure: Mix 0.4 ml of EDTA with 2 ml blood. Fill the Wintrobe tube up to
upper most mark with the help of pasture pipette or syringe. Fill the another Wintrobe tube to balance
first one. If the blood sample is not available, fill the tube with water.

Place the Wintrobe tube in opposite side in centrifuge. Turn the centrifuge to slow speed, then slowly
increase the speed to 3,000 rpm. Centrifuge for 30 min. at 3,000 rpm. After 30 min. switch off the
centrifuge and allow it to stop by itself. Take out the Wintrobe tube and read PCV directly with the
help of graduation mark given on the tube.

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 Microhematocrit Procedure : Draw the blood sample into appropriate capillary tube
with capillary action. Use plain tube for anti-coagulated blood and heparinised tube
for plain blood. In case of finger puncture, the blood should flow freely with little
pressure. Now wipe off the first drop and then collect the blood specimen.

Fill the tube about 3/4th length with blood. Seal another end of the tube with clay or
wax or ultimately by heating. The sealing should be about 2 mm deep. Place two hct
tubes in the groove of centrifuge exactly opposite to each other.

It is not necessary that the capillary tube have exact amount of blood level. In case,
if there is no filled capillary tube to balance, we can use an empty capillary tube.
Centrifuge at 13,000 ± 2000 rpm.

Remove capillary tube from centrifuge. It will show three layers. Top layer is of
plasma or serum; the middle layer is thin creamy white in color and is known as
Buffy coat. It is a layer of WBC; the last layer is the column of RBC. Use the hct
reader for finding the value of hct.

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Aim and principle
Aim of hematocrit or PCV is determine the packed cell volume of the given sample of blood.

The word hematocrit means to separate the blood where the plasma and blood cells are
separated. When a known volume of blood is centrifuged, the cells being heavier, settle down
leaving a clear .This test gives information about RBC concentration and helpful to see
hemoconcentration. And basically, a measurement of total blood volume and RBCs ratio as a
percentage.

Material and reagent

 Macro hematocrit requirement:

1. Blood specimen:
EDTA or double oxalated anti-coagulated blood is used in this method. Determine P.C.V. within
six hr. of blood collection.

2. Wintrobe Tube:
It is 110 mm in length and 2.5 mm in diameter. The lower 100 mm are graduated or marked,
from 100 at top and 0 (zero) at bottom for PCV.

3. Long necked pasture pipette or a special type of syringes is used for filling Wintrobe tube.

4. Centrifuge machine with known speed.

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 Microhematocrit requirement:

1. Blood Specimen:
Blood from finger puncture may be used or EDTA or double oxalate venous blood can also be
used.

2. Capillary Tube:
Use plain capillary tube for anti-coagulated venous blood and use heparinised capillary tube
(Coated with heparin internally) for blood obtained from finger puncture. The capillary tube is
approximately 75 mm in length.

3. Microhematocrit Centrifuge:
This is a special type of centrifuge. It has speed about 15,000 r.p.m. The top of centrifuge is flat
with grooves. The centrifuge also has timer, which is usually set for 5 min.

4. Hematocrit Reader:
There are several types of readers used for reading hct. The simplest method is use of card
reader, which can be made by hand.

5. Clay:
This is used to seal the end of capillary tube.

Normal value and clinical manifestation

Results from your hematocrit test are reported as the percentage of blood cells that are red
blood cells. Normal ranges vary substantially with race, age and sex. The definition of normal
red-blood cell percentage also varies from one medical practice to another.
Generally, a normal range is considered to be:

 For men, 38.3 to 48.6 percent

 For women, 35.5 to 44.9 percent

Children ages 15 and under have a separate set of ranges, as their hematocrit levels change
rapidly with age. The specific lab that analyzes the results will determine the normal hematocrit
range for a child of a certain age.

If your hematocrit levels are too low or too high, it can indicate various problems.

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A lower than normal hematocrit can indicate:

 An insufficient supply of healthy red blood cells (anemia)

 A large number of white blood cells due to long-term illness, infection or a white blood
cell disorder such as leukemia or lymphoma

 Vitamin or mineral deficiencies

 Recent or long-term blood loss

A higher than normal hematocrit can indicate:

 Dehydration

 A disorder, such as polycythemia vera, that causes your body to produce too many red
blood cells

 Lung or heart disease

Discussion
When hematocrit is used to assess anemia and the oxygen-carrying capacity, the advantages and
disadvantages of each method must be carefully considered. In addition, the specific clinical and
analytical needs for any patient population must be determined.

Hematocrit is traditionally a routine hematology laboratory parameter; however, measuring hematocrit as

a STAT parameter in a POC setting is in many ways preferable, as this will save time in critical situations
and avoid specimen transport problems. Some blood gas analyzers offer this possibility, either by
measuring hematocrit by conductivity or by calculating hematocrit from ctHb.

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References:

 https://acutecaretesting.org/en/articles/hematocrit--a-review-of-different-analytical-methods
 https://www.shareyouressays.com/knowledge/method-of-determination-of-packed-cell-volume-
pcv/115433
 PACKED CELL VOLUME OR HAEMATOCRIT
 https://www.getbodysmart.com/circulatory-system/hematocrit-test
 https://www.mayoclinic.org/tests-procedures/hematocrit/about/pac-20384728
 https://www.healthline.com/health/hematocrit#normal-results