Neutrophil Purity
Neutrophil Purity - Related Videos
04:21
Generating Anti-Angiogenic Tumor-Associated Neutrophils
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2025
This video demonstrates an assay to generate anti-angiogenic tumor-associated neutrophils (TANs). Upon taking a murine melanoma tumor containing pro-angiogenic TANs, the single cells are isolated using fluorescence-activated cell sorting (FACS) and treated with an inhibitor of nicotinamide phosphoribosyl transferase (NAMPT) to reduce the capacity to stimulate angiogenesis.
07:12
Neutrophil Isolation Protocol
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Cited by 169 •
2008
Neutrophils are among the first cells to arrive on the site of inflammatory immune response, and their functions and mechanisms have been studied extensively in vitro. We demonstrate a standard density gradient separation method to isolate human neutrophils from whole blood using commercially available separation media.
04:32
Induction and Visualization of Neutrophil Extracellular Traps in Pre-Labeled Neutrophils
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2025
This video demonstrates the induction and visualization of neutrophil extracellular traps or NETs in pre-labeled neutrophils using patient serum containing anti-neutrophil cytoplasmic antibodies. Furthermore, using an impermeable DNA dye and fluorescence microscopy enables the visualization of NETs.
12:17
Detection and Isolation of Apoptotic Bodies to High Purity
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Cited by 35 •
2018
A workflow using flow cytometry or differential centrifugation is developed to detect, quantify and isolate apoptotic bodies from an apoptotic sample to high purity.
12:06
Synthesis of High Purity Nonsymmetric Dialkylphosphinic Acid Extractants
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Cited by 6 •
2017
A protocol for the synthesis of high purity nonsymmetric dialkylphosphinic acid extractants is presented, taking (2,3-dimethylbutyl)(2,4,4'-trimethylpentyl)phosphinic acid as an example.
FAQs
CD16 and CD66b serve as cell-surface markers that help identify neutrophils during sample analysis. Researchers can use their marker profiles in flow cytometry to distinguish the target population from other leukocytes present after isolation. This approach provides a measurable basis for detecting cellular contamination and determining whether the preparation is suitable for neutrophil-focused experiments.
Both density-gradient centrifugation and immunomagnetic separation can enrich neutrophils from blood, but they rely on different separation approaches. Density-gradient centrifugation separates cellular populations during centrifugation, whereas immunomagnetic separation uses magnetic selection to isolate a desired population. Comparing purity after either method helps researchers determine which approach produces a preparation appropriate for a particular assay.
Purity alone does not show whether the isolated neutrophils remain viable. Combining a purity assessment with viability measurements helps distinguish a sample that contains the intended cells from one that also preserves their experimental usefulness. This combined evaluation is important because compromised viability or contaminating leukocytes can alter measured cellular responses and reduce confidence in assay results.
A typical workflow begins by enriching neutrophils from blood using density-gradient centrifugation or immunomagnetic separation. The resulting preparation is then examined by flow cytometry, using markers such as CD16 and CD66b to identify neutrophils and detect contaminating leukocytes. Viability measurements add a second quality check before the sample is used in downstream biological experiments.
Flow cytometry allows researchers to examine marker-defined cell populations rather than treating the preparation as uniformly composed of neutrophils. CD16 and CD66b help identify the target cells, while the observed population can be evaluated for contaminating leukocytes. Adding viability measurements clarifies whether an apparently enriched sample contains cells in a condition appropriate for interpretation.
Purity is particularly important when experiments measure neutrophil-specific antimicrobial activity, inflammation, innate immune responses, or cell signaling. Contaminating leukocytes may contribute their own responses and obscure effects attributed to neutrophils. Assessing the preparation before analysis therefore supports more reliable interpretation, improves comparison across experiments, and helps explain differences in biological assay outcomes.