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Related Experiment Videos

Filipin as a flow microfluorometry probe for cellular cholesterol.

C P Muller, D A Stephany, D F Winkler

    Cytometry
    |January 1, 1984
    PubMed
    Summary
    This summary is machine-generated.

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    Filipin, a fluorescent probe, effectively measures unesterified cholesterol in cells using flow microfluorometry (FMF). This method offers high sensitivity for analyzing cellular cholesterol levels.

    Area of Science:

    • Biochemistry
    • Cell Biology
    • Analytical Chemistry

    Background:

    • Filipin is a polyene antibiotic known to complex with 3 beta-hydroxysterols.
    • Its spectral properties suggest potential for flow microfluorometry (FMF) applications.
    • Accurate measurement of unesterified cellular cholesterol is crucial in biological research.

    Purpose of the Study:

    • To evaluate the efficacy of filipin as an FMF probe for quantifying unesterified cellular cholesterol.
    • To establish optimal experimental conditions for filipin-based FMF analysis.
    • To determine the sensitivity and specificity of filipin for cellular cholesterol detection.

    Main Methods:

    • Utilized fixed murine cells stained with filipin under specific excitation and emission wavelengths.

    Related Experiment Videos

  • Employed flow microfluorometry (FMF) for fluorescence intensity measurements.
  • Validated filipin-based measurements against an independent enzymatic assay for unesterified cholesterol.

    • Main Results:

    • Optimal conditions involved fixed cells, specific filipin concentration, and excitation/emission settings.
    • Filipin-stained cells exhibited significantly increased fluorescence compared to unstained controls.
    • Increased fluorescence intensity strongly correlated with cellular unesterified cholesterol content (P <= .001).
    • The FMF assay demonstrated femtomole sensitivity and distinguished cell populations with varying cholesterol levels.

    Conclusions:

    • Filipin is a valuable and sensitive tool for FMF analysis of unesterified cellular cholesterol.
    • The method allows for the relative quantification of cholesterol in different cell populations.
    • Filipin-based FMF provides a reliable approach for studying cellular sterol metabolism.