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Related Experiment Videos

Flow cytometric reticulocyte analysis. Multiinstitutional interlaboratory correlation study

B H Davis1, N C Bigelow, J A Koepke

  • 1Department of Pathology, Dartmouth Hitchcock Medical Center, Hanover, New Hampshire.

American Journal of Clinical Pathology
|October 1, 1994
PubMed
Summary
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Flow cytometry offers precise reticulocyte analysis, surpassing manual methods. Standardizing this technique, especially with automated analyzers, can reduce interlaboratory variability for reticulocyte counts and maturity index.

Area of Science:

  • Hematology
  • Clinical Pathology
  • Flow Cytometry

Background:

  • Flow cytometry provides superior precision and sensitivity for reticulocyte analysis compared to traditional morphologic methods.
  • It enables the calculation of a reticulocyte maturity index, offering deeper insights into red blood cell production.
  • Previous reports have not extensively detailed interlaboratory variability in flow cytometric reticulocyte analysis.

Purpose of the Study:

  • To evaluate intermethod and interlaboratory variability in flow cytometric reticulocyte analysis.
  • To compare the performance of different flow cytometers and staining methods.
  • To assess the precision of the reticulocyte maturity index across various sites and instruments.

Main Methods:

  • Analysis of 310 samples across eight laboratories using 11 different instruments over a four-month period.

Related Experiment Videos

  • Utilized stains including thiazole orange, ethidium bromide, and auramine O.
  • Employed instruments from Coulter, Becton Dickinson, TOA Medical Electronics, and Ortho Diagnostics.

    • Main Results:

    • The coefficient of variation (CV) for reticulocyte counts varied significantly with reticulocyte percentage, from 69% ( < 0.5% reticulocytes) to 24.1% ( > 2.5% reticulocytes).
    • TOA R-1000 dedicated analyzers demonstrated superior performance with mean CVs of 18.4% ( < 0.5%) and 4.6% ( > 2.5%).
    • The reticulocyte maturity index showed good intersite precision, with mean CVs of 16% (multipurpose cytometers) and 7.3% (TOA R-1000).

    Conclusions:

    • Flow cytometric reticulocyte analysis is demonstrably more precise than manual methods.
    • Greater automation of flow cytometry for reticulocyte analysis can lead to improved interlaboratory standardization.
    • Standardization of reticulocyte counts and maturity index is achievable with further methodological refinement and automation.