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BACKGROUND
The megakaryocytes are large bone marrow cells with a lobulated nucleus and are responsible for the production of thrombocytes. They are produced mainly by the liver, spleen, kidney and bone marrow. They are usually 1 in 10000 bone marrow cells but they can increase in number in certain diseases. They are 10-15 times larger than the red blood cells and measure about 50-100 µm in diameter. The megakaryocyte maturation happens through endomitosis and the nucleus is large and lobulated giving a false impression of multiple nuclei. The nucleus contains 64N DNA or 32 copies of normal DNA. The cytoplasm contains the α granules and dense bodies.1
The aim of the experiment is measurement of diameter of megakaryocyte.
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REQUIREMENTS
Subjects : Human
Apparatus: Ventana Benchmark XT
i View DAB v3 Detection kit
Arcturus XT Laser capture Microdissection System
Haematoxylin and Eosin stains
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PROCEDURE
The specimen is stained with Haematoxylin and Eosin stains for cellular evaluation. The immune histochemical analysis is done by cutting the bone marrow clot and core slides at 4 µm and they are deparaffinized on the Ventana Benchmark XT. The antibody used is mouse anti-CD61 which is incubated for 32 minutes at 37ºC. The antigen was retrieved using the standard cell conditioning solution for 60 minutes using the i View DAB v3 Detection kit. The haematoxylin stain is used to counter stain the slides for 4 minutes. The diameter of the megakaryocyte is measured using the calibrated measurement tool for 400 magnifications using Arcturus XT Laser capture Micro dissection System. The diameter of all the megakaryocytes is measured.2
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CONCLUSION
The size and the number of megakaryocytes help in diagnosis of thrombocytopenia, myelodysplastic syndromes and malignant lymphoma.
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REFERENCES
- Available: https://en.m.wikipedia.org/wiki/Megakaryocyte. Accessed on 11April 2018.
- Fuchs DA, McGinn SG, Cantu CL, Klei RR, Sola-Visner MC, Rimsza LM. Developmental Differences in Megakaryocyte Size in Infants and Children. Am J Clin Pathol. 2012; 138: 140-145.
