Determination of Platelet Count

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BACKGROUND

The platelets also called as thrombocytes play a main role in the stoppage of bleeding along with the coagulation factors in the blood. They do not possess any nucleus with fragments of cytoplasm and are derived from the megakaryocytes. Their main function is coagulation of blood at the site of injury. They follow adhesion, activation, aggregation which activates the coagulation cascade resulting in fibrin deposition and finally form a clot and stop the bleeding.¹

The aim of the experiment is determination of platelet count.

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REQUIREMENTS

Subjects: Human blood sample

Apparatus: Hemocytometer

Unopette capillary pipet, 20 µl capacity

Hemocytometer and coverslip

Microscope

Alcohol pads

Hand counter

Petri dish with moist filter paper

Unopette reservoir no 5854/5855 (1.98 ml of diluent)

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PROCEDURE

Manual platelet count

The blood sample is collected from the subject. Use the protective shield on capillary pipette, puncture diaphragm of Unopette reservoir and add the sample using capillary pipette. Fill the pipet with capillary blood and wipe the extra blood. Mix the blood properly with diluent and let the mixture stand for 10 minutes before charging hemocytometer.

Clean the hemocytometer with alcohol and allow it to dry and charge it. Place the hemocytometer in moist petri dish for 10 minutes and allow the platelets to settle down. Mount the hemocytometer on the microscope and lower its condenser.

Scan the even distribution under 40x magnification and platelets are counted in all 25 small squares within larger center square. Counting has to be done starting from upper left of large middle square. Right hand square, drop down to next row, continue till all 25 squares are completed. Count all the cells that touch upper and left lines and leave out the cells that touch lower or right line.

Platelets /mm³ =average number of platelets × 1000.

Multiply this with 10³ to have platelet count in SI units or 10³/µl.²

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CONCLUSION

The determination of platelet count is important in diagnosis of bleeding disorder, bone marrow disease or any other condition.

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REFERENCES

https://en.m.wikepedia.org/wiki/platelet. Accessed on 05 April 2018.
https://www.austincc.edu/mlt/hem/lab2manplatelet.PDF. Accessed on 05 April 2018.