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Reticulocyte counting using flow cytometry.

P R Nobes1, A B Carter

  • 1Department of Haematology, Middlesex Hospital, London.

Journal of Clinical Pathology
|August 1, 1990
PubMed
Summary
This summary is machine-generated.

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This study refines flow cytometry for routine reticulocyte counting, offering a reliable method for normal and elevated counts. However, it is less accurate for quantifying subnormal reticulocyte levels.

Area of Science:

  • Hematology
  • Clinical Pathology
  • Biotechnology

Background:

  • Accurate reticulocyte quantitation is crucial for assessing bone marrow erythropoietic activity.
  • Traditional methods like visual counting can be labor-intensive and subjective.
  • Flow cytometry offers a potential for automated, objective, and high-throughput analysis.

Purpose of the Study:

  • To refine and validate a flow cytometric method for routine quantitation of reticulocytes.
  • To establish a reference range for the developed method in healthy adults.
  • To compare the flow cytometric method with traditional visual counting techniques.

Main Methods:

  • A flow cytometric assay using thiazole orange staining was developed.
  • Red blood cells were gated based on light scatter properties.

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  • The method was validated against visual counting and tested on patient samples, including those undergoing chemotherapy.

    • Main Results:

    • The flow cytometric method demonstrated high correlation (0.99) with visual counting.
    • A reference range of 19.4-59.2 x 10(9)/l (0.2-1.6%) was established for adults.
    • The method showed good precision for normal and elevated reticulocyte counts but was unreliable for subnormal levels.

    Conclusions:

    • Flow cytometry provides a simple, reliable, and automated method for quantifying normal to high reticulocyte counts.
    • The technique is suitable for routine laboratory use, enhancing diagnostic efficiency.
    • Limitations exist for accurately quantifying very low reticulocyte counts, necessitating caution in specific clinical scenarios.