Platelet Counting by the RBC/Platelet Ratio Method: A Reference Method

  • Klee G
  • D'Onofrio G
  • Van Assendelft O
 et al. 
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Abstract

The International Council for Standardization in Haematology (ICSH) and the International Society of Laboratory Hematology (ISLH) recommend the counting of specifically labeled platelets relative to the RBCs with a fluorescence flow cytometer, together with an accurate RBC count determined with a semiautomated, single-channel aperture-impedance counter as a reference method for the enumeration of platelets. Fresh EDTA-anticoagulated venous blood specimens are measured within 4 hours of the draw. The specimen is prediluted (1:20) and the platelets labeled with two monoclonal antibodies specific to a cluster of differentiation common to all platelets. A final 1:1,000 dilution is made and at least 50,000 events with a minimum of 1,000 platelet events are counted with a flow cytometer to determine the RBC/platelet ratio. The platelet count is then calculated from this ratio and the RBC concentration of the original blood specimen.

Author-supplied keywords

  • Flow cytometry
  • Platelet count
  • Platelets
  • Reference method
  • Value assignment
  • Whole blood calibration

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Authors

  • George Klee

  • Guiseppe D'Onofrio

  • Onno W. Van Assendelft

  • Brian Bull

  • Ahnond Bunyaratvej

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