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Cell surface antigen identification by a modified fluorescein immunosphere method.

J E Tomaszewski, D B Goodman, C M Zmijewski

    American Journal of Clinical Pathology
    |February 1, 1986
    PubMed
    Summary
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    Fluorescein immunospheres offer a viable method for labeling lymphocyte antigens. Stabilizing the antibody-immunosphere complex with glutaraldehyde improves accuracy, rivaling flow cytometry for cell analysis.

    Area of Science:

    • Immunology
    • Cell Biology
    • Biotechnology

    Background:

    • Fluorescein immunospheres are utilized for labeling lymphocyte cell surface antigens via indirect techniques.
    • Wright's counter-staining enables simultaneous assessment of cell morphology and surface antigen characteristics.

    Purpose of the Study:

    • To evaluate methods for separating bound and unbound fluorescein immunospheres from lymphocytes.
    • To optimize the fluorescein immunosphere technique for reliable cell surface antigen analysis.

    Main Methods:

    • Investigated flotation over cold protein solution for unbound immunosphere removal.
    • Employed glutaraldehyde cross-linking to stabilize the antibody-immunosphere-lymphocyte complex.
    • Compared the modified fluorescein immunosphere method with flow cytometry.

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    Main Results:

    • Removal of unbound immunospheres by flotation resulted in significant false negatives.
    • Glutaraldehyde cross-linking effectively stabilized the immunosphere-lymphocyte complex.
    • The modified fluorescein immunosphere method demonstrated performance comparable to flow cytometry.

    Conclusions:

    • Flotation is not a suitable method for removing unbound immunospheres due to false negatives.
    • Glutaraldehyde cross-linking is a key modification for enhancing the reliability of the fluorescein immunosphere technique.
    • This optimized method provides a valuable alternative to flow cytometry for lymphocyte antigen analysis.