Measuring Oxidative Stress in Sepsis

As previously described, a modified procedure was used to measure the generation of reactive oxygen species (ROS) using flow cytometry (21 (link)). Briefly, at 24 hours post-CLP, peritoneal cells were collected. Following RBC lysis, peritoneal cells were loaded in the dark for 30 minutes at 37°C with 5 μM/mL of ROS Brite™ (10 µM; AAT Bioquest, Sunnyvale, CA, USA), which is a general oxidative stress indicator. As soon as the cells were washed and labeled for neutrophil-specific surface markers, flow cytometry analysis was done.

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Zhang T., Zhao J., Fu J., Chen G, & Ma T. (2022). Improvement of the sepsis survival rate by adenosine 2a receptor antagonists depends on immune regulatory functions of regulatory T-cells. Frontiers in Immunology, 13, 996446.

Publication 2022

ROS Brite™

Cells Flow cytometry Neutrophil Oxidative stress Peritoneal Reactive oxygen species

Corresponding Organization : Tianjin Medical University General Hospital

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Variable analysis

independent variables

Time post-CLP (24 hours)

dependent variables

Generation of reactive oxygen species (ROS)

control variables

Incubation time of peritoneal cells with ROS Brite™ (30 minutes)
Concentration of ROS Brite™ (5 μM/mL)
Temperature of incubation (37°C)
Procedures for RBC lysis and labeling of neutrophil-specific surface markers

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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