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    Exercise 5 Hematocrit Determination

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    Jam Ramos
    Here is a labeled illustration of a Wintrobe tube: [ILLUSTRATION OF A LABELED WINTROBE TUBE] The Wintrobe tube has markings along the right side indicating milliliters. It also has a bulb at the bottom and a long neck for centrifugation. Here is an illustration of a packed cell column/volume: [ILLUSTRATION OF A HEMATOCRIT TUBE AFTER CENTRIFUGATION SHOWING THE PACKED RED BLOOD CELLS AT THE BOTTOM] The red blood cells are tightly packed at the bottom of the tube after centrifugation. The buffy coat layer containing white blood cells and platelets

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    Exercise 5 Hematocrit Determination

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    Jam Ramos
    492 views7 pages
    Here is a labeled illustration of a Wintrobe tube: [ILLUSTRATION OF A LABELED WINTROBE TUBE] The Wintrobe tube has markings along the right side indicating milliliters. It also has a bulb at the bottom and a long neck for centrifugation. Here is an illustration of a packed cell column/volume: [ILLUSTRATION OF A HEMATOCRIT TUBE AFTER CENTRIFUGATION SHOWING THE PACKED RED BLOOD CELLS AT THE BOTTOM] The red blood cells are tightly packed at the bottom of the tube after centrifugation. The buffy coat layer containing white blood cells and platelets

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    Here is a labeled illustration of a Wintrobe tube: [ILLUSTRATION OF A LABELED WINTROBE TUBE] The Wintrobe tube has markings along the right side indicating milliliters. It also has a bulb at the bottom and a long neck for centrifugation. Here is an illustration of a packed cell column/volume: [ILLUSTRATION OF A HEMATOCRIT TUBE AFTER CENTRIFUGATION SHOWING THE PACKED RED BLOOD CELLS AT THE BOTTOM] The red blood cells are tightly packed at the bottom of the tube after centrifugation. The buffy coat layer containing white blood cells and platelets

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
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    492 views7 pages

    Exercise 5 Hematocrit Determination

    Uploaded by

    Jam Ramos
    Here is a labeled illustration of a Wintrobe tube: [ILLUSTRATION OF A LABELED WINTROBE TUBE] The Wintrobe tube has markings along the right side indicating milliliters. It also has a bulb at the bottom and a long neck for centrifugation. Here is an illustration of a packed cell column/volume: [ILLUSTRATION OF A HEMATOCRIT TUBE AFTER CENTRIFUGATION SHOWING THE PACKED RED BLOOD CELLS AT THE BOTTOM] The red blood cells are tightly packed at the bottom of the tube after centrifugation. The buffy coat layer containing white blood cells and platelets

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    ROUTINE Hematology LabORATORY

    STUDENT MANUAL ; Volume 1


    Class No. Score

    Name: Avan Dave Mora BSMT : III


    Hennessy Napat-A
    Jam Creibelle Ramos
    Date: October 24, 2021 Section / SET: Set 2-B

    EXERCISE No. 8
    HEMATOCRIT DETERMINATION

    Objectives: The student will be able to:


    1. Determine the different methods of hematocrit determination .
    2. Comprehend the significance and uses of hematocrit.
    3. Prepare a microhematocrit and a macrohematocrit sample.
    4. Perform the hematocrit procedure.
    5. Read and interpret the result obtained from the actual procedures.

    The microhematocrit is a commonly performed test. It may be ordered


    separately or as a part of a Complete Blood Count (CBC). It is a simple
    procedure requiring only two to three drops of blood, which makes it an ideal test
    to follow the progress of anemic or bleeding patients.

    The microhematocrit is a variation of a test called the Hematocrit. The


    hematocrit is a test that is performed using one milliliter of blood in a Wintrobe
    tube. The test is based on the principle of separating the cellular elements of the
    blood from the liquid part, the plasma. In both hematocrit and microhematocrit
    procedures, the separation process is speeded up by centrifugation. After
    centrifugation, the red cells will be at the bottom of the tube, the white cells and
    platelets in the center, and the plasma at the top. The layer containing the white
    cells and platelets has a whitish-tan appearance and is commonly referred to as
    the buffy coat. From this separation the hematocrit or microhematocrit is
    determined by comparing the concentration of the red cells to the volume of the
    whole blood sample. Laboratory personnel often refer to a hematocrit as a “crit”
    or abbreviate it with the letters “Hct”.

    A. ADAM’S MICRO Method

    Materials:
    1. Heparinized capillary tube
    2. Sealing clay
    ROUTINE Hematology LabORATORY
    STUDENT MANUAL ; Volume 1
    3. Hematocrit centrifuge
    4. 70% Alcohol
    5. Cotton
    6. Sterile blood lancets

    Procedure:
    1. Draw blood into a capillet (at least ¾ full) by placing the end of the capillary
    tube over the punctured finger and allow to flow by gravity.
    2. Seal the fine-polished (colored) end of the tube with sealing clay.
    3. Insert the unsealed end of the capillary tube into one end of tube slots near
    the drive shaft of the centrifuge. Then drop the sealed slot into the
    corresponding rim slot.
    4. With the carrying tray in place, the head over the carrying tray. Tighten the
    head cover by turning it by hand in a clockwise direction. Close the
    centrifuge cover.
    5. Turn the centrifuge timer knob to 5 and then turn the switch back to the
    desired running time.
    Note: Centrifuge is done for 5 minutes at
    10,000 revolutions per minute or for 10
    minutes at 5,000 revolutions per minute.
    6. Allow the centrifuge to stop on its own.
    7. Read the value in a microhematocrit reader

    Precautions:
     The recommended speed and time of centrifugation must be strictly
    followed.
     The clay seal must be tight or the contents of the tube may leak out.
     The sealed end of the capillary tube must be placed against the rubber
    gasket in the centrifuge.
     The inner centrifuge lid must be closed securely before closing the outer lid
    to prevent breaking glass tubes.
     The microhematocrit should be read at the top of the red cell layer – not at
    the top of the buffy coat.
    ROUTINE Hematology LabORATORY
    STUDENT MANUAL ; Volume 1

    B. WINTROBE Method:

    Materials
    1. Winthrobe tube
    2. Winthrobe rack
    3. Oxalated blood
    4. Macrocentrifuge

    Procedure
    1. The tube is filled with blood up to the 10th mark of the right side of the tube with
    the use of capillary pipet. Avoid bubbles.
    2. Centrifuge the tube for 30 minutes at 3,500 rpm using the macrocentrifuge.
    (Tube which has been used for ESR determination can be used by just
    centrifuging the tube).
    3. Hematocrit is computed as:

    Ht of packed red blood cells


    Hct (Vol.%) = _________________________________ x 100
    Ht of whole blood used

    Questions:
    1. What are the possible errors in performing hematocrit determination?

    Hematocrit is a blood test done to measure the red blood cells in a


    person’s blood. A high or low RBC count is indicative of a medical
    condition or a disease. In performing the hematocrit determination, few
    errors be may be encountered that could loud to erroneous results. These
    are as follows:

     Excess anticoagulant – when excess amount of anticoagulant is


    added and mixed with the blood, the red cells are destroyed or
    distorted, causing decrease in the hematocrit.
     Insufficient centrifugation – when the hematocrit tubes are
    centrifuged for a short time, there is a false increase in the level of
    hematocrit. The red cells need to settle down at the bottom so they
    need to be centrifuged properly.
    ROUTINE Hematology LabORATORY
    STUDENT MANUAL ; Volume 1
     Inclusion of buffy coat – reading hematocrit should not include the
    buffy coat. Reading should start at the bottom of the buffy coat.
    Inclusion of buffy coat in reading results caused false increase.
     Improper sealing of capillet – this causes false decrease in the
    level of hematocrit. Thick sealing clay should be avoided or more
    than 6 mm.
     Prolonged centrifugation – hematocrit should not be centrifuged
    for more than 5 minutes. This caused too much compaction in the
    red cells causing false decrease in level.
     Dehydration – decreased volume of water causes high hemoglobin
    thus causing false increase in the level of hematocrit. The proportion
    of water to the formed elements is not relative.

    2. What are the advantages of Micro Hematocrit Method?

    There are few advantages of microhematocrit methods over other


    methods. In performing microhematocrit procedures, only small amount of
    blood is required so venipuncture may be avoided. The procedure is also
    easy to perform and provide faster results. No dilution is needed in this
    method and hemolysis can be detected when reading the results.

    3. What are the other methods of Hematocrit determination?

    There are various methods for hematocrit determination and these are
    categorized as micromethods and macromethods.

    For macromethods:
    A. Wintrobe method that uses double oxalate as an
    anticoagulant.
    B. Haden’s modification method that uses sodium oxalate as an
    anticoagulant.
    C. Van Allen’s method that uses sodium oxalate as an
    anticoagulant.
    D. Sanfor-Magath method that uses sodium oxalate as an
    anticoagulant.
    E. Bray’s method that uses heparin as an anticoagulant.

    For micromethod:
    A. Adam’s method that uses heparinized capillary hematocrit
    tube.
    ROUTINE Hematology LabORATORY
    STUDENT MANUAL ; Volume 1

    4. What are the uses of buffy coat?

    A buffy coat is a concentrated suspension of WBCs and platelets that make


    up a part of the anticoagulated blood sample obtained by centrifugation. This
    is used on various purposes, such as but not limited to:
     DNA isolation from blood samples.
     It helps concentrate large volumes of blood samples so that it
    decreases the downstream during cell separation and handling.
     It reduces donor variability.
     It improves platelet yield.
     It helps detect infection with malaria and blood parasites.

    5. What is the clinical significance of determining the packed cell volume?

    Hematocrit test is used to check the proportion of red blood cells. Determining
    the packed cell volume is very important, because it provides a rough
    observation and estimation of the size and concentration of the red blood cells. It
    aids in the diagnosis of disorders such as anemia, leukemia, polycythemia and
    dehydration. It also helps monitors if the body is responding to certain
    treatments.
    ROUTINE Hematology LabORATORY
    STUDENT MANUAL ; Volume 1
    ==============
    ILLUSTRATIONS:
    ==============
    a. Draw and label a Wintrobe tube
    ROUTINE Hematology LabORATORY
    STUDENT MANUAL ; Volume 1
    ==============
    ILLUSTRATIONS:
    ==============
    b. Illustrate a packed cell column/volume.

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