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Related Experiment Videos

Quantitation of cell concentration using the flow cytometer

C C Stewart, J A Steinkamp

    Cytometry
    |January 1, 1982
    PubMed
    Summary
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    A new flow cytometry method accurately counts cells and analyzes marker-positive populations. This technique uses fluorescent particles to determine sample volume, enabling precise cell concentration calculations, even in complex samples like tumors.

    Area of Science:

    • Biotechnology
    • Cell Biology
    • Analytical Chemistry

    Background:

    • Accurate cell concentration measurement is crucial for biological research and diagnostics.
    • Traditional methods can be time-consuming and less precise, especially with complex samples.
    • Flow cytometry offers high-throughput analysis but requires robust quantification techniques.

    Purpose of the Study:

    • To develop a simple, simultaneous method for measuring cell concentration and analyzing marker-positive cell populations using flow cytometry.
    • To establish a reliable quantification strategy applicable to challenging biological samples.

    Main Methods:

    • Incorporation of fluorescent latex particles of known concentration into cell samples.
    • Analysis of both marker-positive cells and fluorescent particles via flow cytometry.

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  • Calculation of sample volume based on accumulated particle count.
  • Determination of total and marker-positive cell concentrations.

    • Main Results:

    • Demonstrated simultaneous measurement of cell concentration and marker-positive cell populations.
    • Validated the method using single, dual, and triple laser excitation with DNA-stained cells and fluorescent spheres.
    • Successfully applied the procedure to count cells from highly necrotic tumors with significant debris.

    Conclusions:

    • The developed flow cytometric procedure provides a simple and accurate method for cell concentration and marker analysis.
    • This technique enhances quantification capabilities, particularly in samples with high levels of cellular debris.
    • The method offers a valuable tool for cell-based assays and biological sample analysis.