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Bone marrow cell differential counts obtained by multidimensional flow cytometry.

L W Terstappen1, J Levin

  • 1Becton Dickinson Immunocytometry Systems, San Jose, California 95131.

Blood Cells
|January 1, 1992
PubMed
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Five-dimensional flow cytometry quantifies normal bone marrow cells, including neutrophils and lymphocytes. This method accurately differentiates cell types and aids in quantitative bone marrow differentials, potentially reducing reliance on manual smear reviews.

Area of Science:

  • Hematology
  • Immunology
  • Biotechnology

Background:

  • Accurate enumeration of bone marrow cell populations is crucial for diagnosing hematological disorders.
  • Traditional methods like light microscopy for bone marrow differentials can be labor-intensive and subjective.
  • Flow cytometry offers a high-throughput, quantitative approach to cellular analysis.

Purpose of the Study:

  • To establish a five-dimensional flow cytometric method for analyzing normal bone marrow aspirates.
  • To determine the frequencies of major nucleated bone marrow cell subpopulations.
  • To validate the technique by comparing flow cytometry results with cell sorting and light microscopy.

Main Methods:

  • Utilized five-dimensional flow cytometry on normal bone marrow aspirates from 50 donors.

Related Experiment Videos

  • Employed specific fluorescent stains (Thiazole-Orange, LDS-751) and antibodies (CD45-PE) for cell identification.
  • Verified cell population identities through cell sorting and subsequent light microscopic examination.
  • Analyzed forward light scatter, orthogonal light scatter, and staining intensity for unique cellular features.

    • Main Results:

    • Established mean frequencies and ranges for neutrophils, eosinophils, monocytes, lymphocytes, nucleated erythrocytes, and a mixed population including blasts, megakaryocytes, plasma cells, and basophils.
    • Demonstrated significantly higher reticulocyte frequencies in bone marrow compared to peripheral blood (p < 0.0001).
    • Successfully discriminated between immature and mature reticulocytes based on differential staining intensity.
    • Observed higher nucleated cell frequencies and lower platelet frequencies in bone marrow aspirates versus peripheral blood.

    Conclusions:

    • Five-dimensional flow cytometry provides a robust and quantitative method for bone marrow cell differential analysis.
    • The technique can accurately identify and quantify various bone marrow cell populations, including immature reticulocytes.
    • This flow cytometric approach has the potential to streamline laboratory workflows and reduce the need for manual microscopic examination of bone marrow smears.