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Related Experiment Videos

A rapid quantitative assay for activated neutrophils

W W Bullock, H R Creamer, W L Gabler

    Inflammation
    |September 1, 1982
    PubMed
    Summary

    A new assay rapidly detects activated human neutrophils (PMNs) by measuring decreased optical density of cell suspensions. This sensitive method allows analysis of 400 samples daily from minimal blood volumes.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Assay Development

    Background:

    • Neutrophil activation is crucial in immune responses.
    • Existing assays for activated neutrophils can be time-consuming or require large sample volumes.
    • A need exists for rapid, sensitive, and high-throughput methods to quantify neutrophil activation.

    Purpose of the Study:

    • To develop and validate a novel, rapid, and sensitive assay for activated human neutrophils.
    • To establish a quantitative method for measuring neutrophil aggregation using optical density.
    • To assess the assay's sensitivity and throughput capacity.

    Main Methods:

    • Developed a quantitative assay measuring the decrease in optical density (OD) of aggregated human neutrophil suspensions.
    • Utilized an ELISA machine for time-dependent OD measurements of activated neutrophils (5 x 10^5 cells/well) in microtiter plates.
    • Tested assay sensitivity using lipopolysaccharide (LPS) and Con-A-activated supernatant.

    Main Results:

    • The assay demonstrated high sensitivity, detecting aggregation induced by 0.0001 µg/ml LPS.
    • It could detect activation by lymphokines in Con-A-activated supernatant diluted 1/500.
    • The assay allows for the analysis of up to 400 neutrophil suspensions per day from less than 80 ml of blood.

    Conclusions:

    • The developed optical density assay is a rapid, sensitive, and convenient method for quantifying activated human neutrophils.
    • This assay offers high throughput, enabling the analysis of numerous samples efficiently.
    • The method provides a valuable tool for immunological research and diagnostics requiring neutrophil activation assessment.

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