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Related Experiment Video

Updated: Oct 29, 2025

Study of Phagolysosome Biogenesis in Live Macrophages
08:06

Study of Phagolysosome Biogenesis in Live Macrophages

Published on: March 10, 2014

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Quantifying phagosomal HOCl at single immune-cell resolution.

Palapuravan Anees1, Matthew Zajac1, Yamuna Krishnan1

  • 1Department of Chemistry, University of Chicago, Chicago, IL, United States; Grossman Institute of Neuroscience, Quantitative Biology and Human Behavior, University of Chicago, Chicago, IL, United States.

Methods in Cell Biology
|July 6, 2021
PubMed
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Researchers developed a novel fluorescent DNA reporter to map hypochlorous acid (HOCl) dynamics within phagosomes. This method provides sub-cellular resolution for studying pathogen killing and immune cell activity.

Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • Phagocytic immune cells neutralize pathogens within phagosomes, a compartment requiring reactive species for killing.
  • Hypochlorous acid (HOCl), a key reactive species, is produced by myeloperoxidase (MPO) within phagosomes.
  • Monitoring HOCl dynamics is crucial for understanding pathogen killing and host-pathogen interactions, but current methods lack spatial resolution.

Purpose of the Study:

  • To develop and validate a novel method for quantifying HOCl dynamics in phagosomes.
  • To enable sub-cellular and single-cell resolution mapping of HOCl production during phagosome maturation.

Main Methods:

  • A fluorescent DNA-based reporter was designed to detect and quantify HOCl.
  • The protocol was optimized for assaying HOCl dynamics in phagosomes with fluctuating pH.
Keywords:
DNA-based reporterFluorescent probeHypochlorous acidImmune cellsMyeloperoxidasePhagosomepH

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Related Experiment Videos

Last Updated: Oct 29, 2025

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  • The reporter's performance was compared to traditional HOCl visualization and MPO activity assays.

    • Main Results:

    • The developed method allows for precise mapping of HOCl within phagosomes.
    • It offers superior sub-cellular spatial resolution compared to existing techniques.
    • The reporter enables HOCl production assays at the single-cell level.

    Conclusions:

    • This novel fluorescent DNA reporter provides a powerful tool for studying HOCl dynamics in phagosomes.
    • The method enhances our ability to investigate immune cell function, pathogen resistance, and host-pathogen interactions.
    • It overcomes limitations of previous methods, enabling detailed analysis of reactive species production in cellular compartments.