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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Analyzing Platelet Subpopulations by Multi-color Flow Cytometry
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A flow cytometric method for platelet counting in platelet concentrates.

Pieter F van der Meer1, Willy Karssing-van Leeuwen, Jim Kurtz

  • 1Sanquin Blood Supply, Amsterdam, The Netherlands. p.vandermeer@sanquin.nl

Transfusion
|July 28, 2011
PubMed
Summary

A new flow cytometric method for platelet (PLT) counting in PLT concentrates offers improved inter-center comparisons. While individual hematology analyzers are more precise, the flow cytometry method serves as a valuable calibration tool.

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Area of Science:

  • Hematology
  • Transfusion Medicine
  • Flow Cytometry

Background:

  • Platelet (PLT) concentrates are counted using hematology analyzers, but results vary significantly between devices, hindering comparisons.
  • The International Council for Standardization in Hematology (ICSH) developed a modified reference method for PLT concentrates due to the absence of red blood cells.

Purpose of the Study:

  • To validate a modified ICSH reference method for platelet counting in platelet concentrates.
  • To compare the performance of a novel flow cytometric method developed by the Biomedical Excellence for Safer Transfusion (BEST) Collaborative against standard hematology analyzers.

Main Methods:

  • Five platelet concentrate samples were distributed to eight participating centers for same-day counting.
  • Platelets were stained with anti-CD41a and analyzed via flow cytometry using a uniform template.
  • Samples were also counted on 15 different hematology analyzers.

Main Results:

  • The BEST flow cytometric method and the modified ICSH method showed less than 1% difference in platelet counting results.
  • The inter-center coefficient of variation (CV) was 6.3% for the BEST method versus 7.6% for hematology analyzers.
  • Individual hematology analyzers demonstrated higher precision (mean CV of 0.9%) compared to flow cytometers (mean CV of 3.7%).

Conclusions:

  • The BEST flow cytometric method provides a smaller inter-center CV and better center-to-center agreement than hematology analyzers.
  • Hematology analyzers are more precise for individual measurements, but the flow cytometric method is valuable for inter-center calibration.
  • Routine platelet counting should continue with hematology analyzers, with the flow cytometric method serving as a calibration standard.