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Updated: Jan 23, 2026
10:33
Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages
Published on: October 25, 2017
11.2K
Kinetic Analysis of Phagosomal ROS Generation
Sophie Dupré-Crochet1, Marie Erard1, Oliver Nüβe2
1LCP, CNRS UMR 8000, Université Paris-Sud, Université Paris-Saclay, Orsay, France.
Methods in Molecular Biology (Clifton, N.J.)
|June 8, 2019
Summary
This study introduces a method to track reactive oxygen species (ROS) production within phagosomes. This technique aids in understanding immune responses against microbes, especially those resisting destruction.
Area of Science:
- Immunology
- Cell Biology
- Microbiology
Background:
- Phagosomal reactive oxygen species (ROS) generation is crucial for immune defense against microbial infections.
- Accurate measurement of phagosomal ROS is needed to understand host-pathogen interactions, particularly with resilient microbes.
- Detecting ROS is challenging due to their short lifespan and reactivity.
Purpose of the Study:
- To develop and validate a method for quantifying phagosomal ROS production kinetics.
- To enable detailed analysis of the interplay between phagocytes and microbes.
- To provide a tool for studying pathogens that evade phagosomal destruction.
Main Methods:
- Directly labeling phagocytic prey with a ROS-sensitive dye.
- Targeting the dye specifically into the phagosome.
- Developing quality assessment and imaging techniques for kinetic analysis.
Main Results:
- The described labeling procedure successfully targets ROS-sensitive dyes into phagosomes.
- The method allows for kinetic analysis of ROS production at the single-phagosome level.
- Quality assessment and imaging techniques were established for reliable data acquisition.
Conclusions:
- Direct labeling of prey offers a viable strategy for studying phagosomal ROS dynamics.
- This approach enhances our understanding of immune cell function during infection.
- The technique is valuable for investigating microbial resistance mechanisms to phagosomal degradation.
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