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.

2022年3月7日 · By comparing eight methods of neutrophil characterization, we demonstrate that the level of neutrophil activation and degranulation is associated with specific experimental conditions and the...

Flow cytometry analysis of neutrophils Under homeostatic conditions, neutrophils are most prevalent circulating in the periphery, making flow cytometry a convenient tool for their characterization. A list of relevant immunophenotyping markers can be found in Table 2 .

Although neutrophils can be visually identified based on the shape of their nuclei and cytoplasmic granularity, they can also be identified by flow cytometry. Surface markers are used to identify and classify subsets of neutrophils. These neutrophil markers are important for activation and cellular function of both mature and immature neutrophils.

Polymorphonuclear neutrophils (PMNs) protect the host from invading microorganisms. However, excessively activated PMNs can also cause damage to host tissues under inflammatory conditions. Here we developed simple assays to determine the activation state of PMNs in human whole blood that contains so …

2021年11月13日 · 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.

1999年12月17日 · This review evaluates the various methods available for measuring neutrophil function using flow cytometry. This powerful tool has the ability to measure multiple parameters on individual cells at high speed.

Here we describe a novel flow cytometry (FLOW)-based assay to identify and quantify NETs using antibodies against key NETs constituents, specifically DNA, modified histones, and granular enzymes. This method is applicable to both murine and human samples for the assessment of induced NETs in vitro, or detection of NETosis in vivo in blood samples.

We developed a high throughput flow cytometry-based assay for detecting changes to four canonical neutrophil functions following biological or chemical triggers. Our assay detects neutrophil phagocytosis, reactive oxygen species (ROS) generation, ectodomain shedding, and secondary granule release in a single reaction mixture.

1999年12月17日 · Flow cytometric analysis provides a rapid screen for abnormalities of neutrophil function and reflects more accurately their behavior in vivo. Neutrophils are important effector cells in numerous inflammatory conditions.