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Related Experiment Videos

High-Throughput Particle Uptake Analysis by Imaging Flow Cytometry.

Asya Smirnov1, Michael D Solga2, Joanne Lannigan1

  • 1Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia.

Current Protocols in Cytometry
|April 4, 2017
PubMed
Summary

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This study introduces a high-throughput imaging flow cytometry method to quantify particle uptake by host cells, exemplified by bacterial internalization in neutrophils. This protocol accurately measures bacterial association and internalization efficiency in thousands of cells.

Area of Science:

  • Cell biology
  • Immunology
  • Microbiology
  • Drug delivery

Background:

  • Quantifying host cell particle uptake is crucial across various biological and medical fields.
  • Existing methods may lack the throughput for comprehensive analysis.
  • Understanding bacterial internalization by immune cells like neutrophils is key in infectious disease research.

Purpose of the Study:

  • To develop and validate a high-throughput protocol for analyzing particle uptake by host cells using imaging flow cytometry.
  • To quantify the efficiency of bacterial association and internalization by neutrophils.
  • To provide a versatile method applicable to diverse experimental systems.

Main Methods:

  • Utilized imaging flow cytometry for high-throughput analysis of particle-host cell interactions.
Keywords:
attachmentbacteriaimaging flow cytometryinternalizationphagocytosis

Related Experiment Videos

  • Employed fluorescently labeled bacteria (Neisseria gonorrhoeae) and a bacteria-specific antibody with a distinct fluorophore.
  • Developed a spot count algorithm to differentiate extracellular (double-labeled) and intracellular (single-labeled) bacteria within individual host cells.

    • Main Results:

    • Successfully quantified bacterial association and internalization percentages in neutrophils.
    • Demonstrated the ability to distinguish between surface-bound and internalized bacteria.
    • The protocol enables analysis across thousands of cells, providing statistically robust data on uptake efficiency.

    Conclusions:

    • The presented imaging flow cytometry protocol offers a robust and high-throughput method for quantifying particle uptake.
    • This technique is valuable for studying host-pathogen interactions, particularly bacterial internalization by phagocytic cells.
    • The protocol's adaptability makes it suitable for various research areas, including infectious diseases, cancer, and drug delivery.