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Automation of the Micronucleus Assay Using Imaging Flow Cytometry and Artificial Intelligence
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Normalized neutral lipid quantitation by flow cytometry.

Nathan E Wolins1, Katerina N DeHaan2, Vincenza Cifarelli1

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|May 17, 2018
PubMed
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Flow cytometry can now quantify cellular lipid levels by measuring fluorescence intensity. This method allows accurate estimation of triacylglycerol mass in mixed cell populations, aiding metabolic disease research.

Keywords:
BODIPY® 493/503lipid dropletmonocytesperilipintriacylglycerol

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Metabolic Disease Research

Background:

  • Measuring tissue lipids is crucial for understanding metabolic diseases.
  • Current methods for assessing cellular lipid levels, like flow cytometry, have limitations in inter-sample comparisons.
  • Linking specific cell types to disease pathogenesis requires precise lipid quantification within heterogeneous tissues.

Purpose of the Study:

  • To develop a flow cytometry-based method for quantifying cellular lipid levels.
  • To enable the estimation of triacylglycerol (TG) mass in mixed cell populations.
  • To overcome limitations of existing techniques for lipid analysis in specific cell types.

Main Methods:

  • Utilizing lipophilic fluorophore staining for flow cytometry analysis.
  • Establishing a calibration curve where fluorescence intensity correlates linearly with lipid levels.
  • Normalizing fluorescence intensity to control conditions to reduce experiment-to-experiment variation.

Main Results:

  • Fluorescence intensity in stained cells accurately reflects cellular lipid content.
  • The method allows for lipid level assessment in mixed cell populations.
  • Triacylglycerol (TG) mass can be estimated in specific cell populations within a mixture.
  • Confirmed perilipin 1 (PLIN1) enhances TG accumulation in Huh7 cells.

Conclusions:

  • Flow cytometry offers a robust method for quantifying cellular lipid levels and estimating TG mass.
  • This technique facilitates lipid analysis in challenging cell populations, such as human blood monocytes.
  • The developed method addresses limitations in biochemical assays for TG in low-cell-number or isolated populations.