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Two-color immunofluorescence using a fluorescence-activated cell sorter.

M R Loken, D R Parks, L A Herzenberg

    The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
    |July 1, 1977
    PubMed
    Summary
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    A novel flow system technique allows simultaneous analysis of fluorescein and rhodamine dyes using single excitation wavelength. This method aids in studying immunoglobulin M and immunoglobulin D on mouse splenic lymphocytes.

    Area of Science:

    • Immunology
    • Biochemistry
    • Analytical Chemistry

    Background:

    • Flow cytometry is crucial for analyzing cell populations.
    • Distinguishing multiple fluorochromes simultaneously can be challenging.
    • Immunoglobulin M (IgM) and Immunoglobulin D (IgD) are key B-cell surface markers.

    Purpose of the Study:

    • To develop a flow cytometry technique for simultaneous analysis of fluorescein and rhodamine.
    • To apply this technique to investigate the expression of IgM and IgD on mouse splenic lymphocytes.

    Main Methods:

    • Utilized a single excitation wavelength for fluorescein and rhodamine analysis.
    • Employed optical and electronic signal discrimination methods.
    • Analyzed mouse splenic lymphocytes using the developed flow system.

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    Main Results:

    • Successfully discriminated fluorescein and rhodamine signals in a flow system.
    • Demonstrated the utility of the technique for studying cell surface markers.
    • Provided insights into the relationship between IgM and IgD expression on lymphocytes.

    Conclusions:

    • The described flow system technique enables efficient simultaneous detection of fluorescein and rhodamine.
    • This method is valuable for immunological studies involving multiple B-cell markers.
    • Further research can explore this technique in various cell types and biological systems.