Protocol for analysis of mouse neutrophil NETosis by flow cytometry

Abstract

Summary: Studies involving neutrophils are steadily increasing, thus creating a need for more optimized and thorough protocols for studying neutrophil function. Here, we present our protocol for extracting mouse bone marrow neutrophils, estimating the purity of isolated neutrophils, and assessing their ability to induce NETosis upon an external cue. We test two isolation protocols that can be used to attain neutrophils to assess NETosis induction. This approach allows for the parallel assessment of NETosis induction in cohorts larger than 10 samples.For complete details on the use and execution of this protocol, please refer to Lu et al., 2021.

Keywords

• Cell isolation
• Flow Cytometry/Mass Cytometry
• Cell-based Assays
• Immunology