MEDT 20: HEMATOLOGY 2 LABORATORY

Platelet Structure and Platelet Estimate

Learning Outcomes

After the completion of this module, you will be able to:

1. Identify platelets
2. Differentiate platelets from other debris
3. Demonstrate how to count platelets using manual method
4. To compute the platelet count and report the results using conventional
and SI units
5. To identify thrombocytosis and thrombocytopenia

Introduction

Platelets are fragments arise from the megakaryocyte, which is the largest cell found in
the bone marrow. (Monocytes, largest in the peripheral blood). This cell undergoes
endomitosis, which is a process of nuclear division without cellular division (Themes 2016).
They are small, colorless, moderately refractile bodies, that appears as azure granules with
scanty light blue cytoplasm when stained (Sadang et al. 2015). Platelets are essential for
maintaining the balance of bleeding and clotting within the body (Hemostasis), by this platelet
count gives a clue for what state a patient is in, or what state the patient will be.

Materials

1. Phlebotomy Kit
2. Anticoagulated Blood(EDTA)
3. Hema Quick-Giemsa stain
4. Glass slide
5. Applicator stick
6. Microscope
7. Personal Protective Equipment

Notes
I. Platelet structure under the microscope

Figure 1. The picture on the left, peripheral blood smear stained by Giemsa stain. The
picture on the right an activated and inactivated platelets under electron microscope.

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o Platelets are around 2-3 um(diameter), that adhere, aggregate and secrete the
contents of their granules

Figure 2. Platelet clumping (left and right)

Figure 3. Platelet satellitism

Figure 4. Giant platelets

II. Platelet estimate

A platelet count is usually part of a complete blood count. This test measures the number
of platelets, white blood cells and red blood cells. An estimated platelet count can be done
by simply preparing a blood smear. Then, it will be evaluated at 100× magnification (OIO).
The platelet count/ uL of blood can be approximated by multiplying the number of platelets
(average) seen in 10 microscope field by 15,000.

Procedure

1. Perform a venipuncture and use the lavender top (EDTA) for collection

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 MEDT 20: HEMATOLOGY 2 LABORATORY

2. Prepare a blood smear

3. Dry the smear

4. Stain the prepared smear (using the manufacturer’s guide for staining)

5. View the smear under the microscope (HPO)

6. Count the number of platelets in ten field under 100x magnification

7. Average the platelet count

8. Multiply the average by 15,000

Report the platelet count

III. Other methods of Platelet Count
a. Reese-Ecker Method
Diluting fluid composition: Brilliant cresyl blue, 40% formalin, and sodium Citrate

Procedure:
1. Draw blood up to 0.5 mark of the RBC pipette
2. Dilute blood with Reese Ecker diluting fluid up to 101 marks.
3. Shake the pipette for 1-5 minutes
4. Discard 5-6 drops and charge the counting chamber
5. Place the counting chamber on a petri dish with a wet filter paper to prevent evaporation.
Let it stand for 10-15 minutes to allow the platelet to settle.
6. Cout the platelets in all 25 tertiary squares of central secondary square (1mm2)
*Platelets are stained light blue

Computation:

No. of cells/mm3=Total no. of cells counted

area (1) x depth (1/10) x dilution (1/200)
or cells counted x 2,000
b. Ammonium Oxalate technique
Procedure:

1. Draw blood up to the 1.0 mark of the RBC pipette and ammonium oxalate up

to the 101 marks.

2. Procedure 2-6 of Reese-Ecker Method perform

Computation:

No. of cells/mm3=Total no. of cells counted

area (1) x depth (1/10) x dilution (1/100)
or cells counted x 1,000

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C. Direct Method
A. Nygard’s Sources of Error:
B. Guys and Leake’s
C. Brenker-Cronkite 1. Platelet clumps
2. Imperfect sources of blood
D. Indirect Method 3. Blood in EDTA kept 20 degrees Celsius
A. Dameshek
B. Fonio’s
C. Olef’s-The best indirect method

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 MEDT 20: HEMATOLOGY 2 LABORATORY
Date
Name/s

Year/Section/Cluster

1. Post the picture of the fields where you count the platelets (10 fields) (10 Points)

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 MEDT 20: HEMATOLOGY 2 LABORATORY

2. Show the computation of the platelet count (5 points)

Platelet count report__________

Name of the Patient________________________

3. What are the conditions that may lead to thrombocytosis and thrombocytopenia?

Thrombocytosis Thrombocytopenia

References:

 Sadang M.M and Llanera, F.R (2015) Laboratory Manual in Hematology

 Elaine M. Keohane, Larry J. Smith, Jeanine M. Walenga 2016 Rodak's
Hematology Clinical Principles and Applications 5th Edition

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