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Related Experiment Videos

Reticulocyte counting by flow cytometry. A comparison with manual methods.

A A Pappas1, R B Owens, J T Flick

  • 1Department of Pathology, University of Arkansas for Medical Sciences, Little Rock 72205.

Annals of Clinical and Laboratory Science
|March 1, 1992
PubMed
Summary
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Fluorescent activated cytometric (FACS) analysis offers a precise and rapid method for reticulocyte counting, improving anemia diagnosis and therapy monitoring. This automated approach overcomes manual method limitations, providing valuable insights into erythropoietic activity.

Area of Science:

  • Hematology
  • Clinical Diagnostics
  • Flow Cytometry

Background:

  • Reticulocyte count (RC) is crucial for anemia evaluation and therapy response.
  • Manual RC methods suffer from imprecision and inter/intraobserver variability.
  • Automated methods offer improved accuracy and speed for reticulocyte enumeration.

Purpose of the Study:

  • To evaluate fluorescent activated cytometric (FACS) analysis for reticulocyte counting.
  • To compare FACS reticulocyte counts with manual methods.
  • To explore the utility of the reticulocyte maturation index (RMI) derived from FACS.

Main Methods:

  • Reticulocytes were analyzed using thiazole orange (TO) staining via FACS.
  • FACS automated counting of over 10,000 cells compared to manual 1,000 cell counts.

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  • Reticulocyte maturation index (RMI) was determined as a measure of RNA content.

    • Main Results:

    • FACS reticulocyte counts demonstrated good correlation with manual methods and similar reference ranges.
    • FACS offers enhanced precision due to higher cell counts.
    • RMI provides an independent measure of erythropoietic activity, potentially aiding in predicting engraftment and subclassifying anemias.

    Conclusions:

    • FACS determination of reticulocyte counts provides significant advantages over manual methods for monitoring erythropoiesis.
    • Awareness of potential pitfalls, such as interfering substances, is necessary for accurate FACS interpretation.
    • The reticulocyte maturation index (RMI) offers a novel parameter for assessing erythropoietic function.