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Related Experiment Videos

Lymphocyte subset determination using a hematology analyzer

J C Hudson1, R F Brunhouse, C Garrison

  • 1Coulter Corporation, Miami, FL 33116-9015, USA.

Cytometry
|June 15, 1995
PubMed
Summary
This summary is machine-generated.

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This study introduces a novel method using anti-CD4 antibody microspheres to identify CD4 cells in blood. The VCS-microsphere technique shows high correlation with traditional flow cytometry, potentially simplifying CD4+ cell analysis.

Area of Science:

  • Immunology
  • Hematology
  • Analytical Chemistry

Background:

  • Accurate enumeration of CD4+ T-lymphocytes is crucial for monitoring immune status, particularly in conditions like HIV/AIDS.
  • Traditional flow cytometry methods for CD4+ cell counting can be complex and require specialized gating strategies.

Purpose of the Study:

  • To evaluate a novel method for quantifying CD4+ lymphocytes using anti-CD4 antibody-coated microspheres and a modified Coulter VCS hematology analyzer.
  • To compare the accuracy of the VCS-microsphere method with conventional flow cytometry for CD4+ cell enumeration.

Main Methods:

  • Whole blood samples were incubated with anti-CD4 antibody-coated microspheres to label CD4+ cells.
  • Cells were lysed and analyzed using a modified Coulter VCS analyzer, differentiating labeled cells by changes in volume, conductance, and light scatter.

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  • Results were compared to standard flow cytometry, with and without CD3 gating.

    • Main Results:

    • The VCS-microsphere method demonstrated a high correlation (R = 0.996) with conventional flow cytometry for CD4+ lymphocyte quantification.
    • A regression analysis showed a slope of 0.96, indicating close agreement between the methods when CD3 gating was applied to flow cytometry data.
    • CD3 gating, while necessary for traditional flow cytometry scatterplots, appeared potentially unnecessary for the VCS-microsphere volume-conductance-light scatterplots.

    Conclusions:

    • The anti-CD4 antibody-coated microsphere method combined with VCS analysis offers a reliable and potentially simplified approach for CD4+ lymphocyte enumeration.
    • This technique may reduce the complexity of CD4+ cell counting, offering a valuable alternative for clinical and research settings.