Neutrophil Monocyte Gating
Neutrophil Monocyte Gating - Related Videos
09:28
Imaging Neutrophils and Monocytes in Mesenteric Veins by Intravital Microscopy on Anaesthetized Mice in Real Time
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Cited by 6 •
2015
We detail a protocol to monitor the behavior of neutrophils and monocytes in mesenteric veins under steady state and inflammatory conditions using intravital confocal microscopy on anaesthetized mice.
12:29
Live Imaging of Antifungal Activity by Human Primary Neutrophils and Monocytes in Response to A. fumigatus
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Cited by 10 •
2017
Here, we describe a protocol to assess antifungal activity of primary human immune cells in real-time using fluorescent Aspergillus reporter conidia in conjunction with live-cell video microscopy and flow cytometry. Generated data provide insight into host cell-Aspergillus interactions such as fungicidal activity, phagocytosis, cell migration and inhibition of fungal growth.
07:12
Neutrophil Isolation Protocol
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Cited by 173 •
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.
11:29
Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood
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Cited by 5 •
2016
The purpose of this manuscript is to present a method for measuring monocyte and granulocyte phagocytosis and oxidative burst activity in human blood samples.
08:31
Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
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Cited by 19 •
2016
This study compares two different methods of human monocyte isolation for obtaining in vitro dendritic cells (DCs). Monocytes are selected by adherence or negatively enriched by magnetic separation. Monocyte yield and viability along with MDDC viability, proliferation and CD11c/CD14 surface marker expression will be compared between both methods.
FAQs
Forward scatter and side scatter add physical information to marker-based identification. These signals help distinguish cellular populations from debris and provide an initial view of differences within a mixed blood or tissue sample. Surface markers then refine the assignment of neutrophil and monocyte populations, improving the consistency of downstream measurements of abundance and immune-state changes.
CD16 can contribute to identifying both neutrophils and monocytes, so it cannot always serve as an isolated discriminator between these populations. Its interpretation should be considered alongside other lineage-associated markers, such as CD66b for neutrophils or CD14 for monocytes, and the physical characteristics of the cells. This combined strategy supports clearer population boundaries.
The two information sources address different sources of uncertainty. Scatter characteristics help identify cellular structure and remove debris, whereas markers provide lineage-associated evidence for assigning cells to neutrophil or monocyte populations. Using both reduces reliance on a single feature and supports reproducible quantification when samples contain multiple leukocyte populations or variable inflammatory cell distributions.
A practical analysis begins by examining the sample’s scatter profile to separate cellular events from debris and other unsuitable material. Lineage-associated markers are then used to define neutrophil and monocyte populations within the retained events. The resulting gates can support population quantification and comparison of immune characteristics across blood or tissue samples.
After the populations are defined, the analysis can measure changes in cell abundance, activation, maturation, and recruitment. These readouts help connect shifts in neutrophil or monocyte populations with the intensity or character of an innate immune response. Comparing gated populations across samples can therefore reveal coordinated changes in inflammatory status rather than cell counts alone.
In infection research, the strategy helps characterize innate immune responses during host-pathogen interactions. Researchers can examine whether neutrophil or monocyte populations change in abundance, activation, maturation, or recruitment, then relate those patterns to inflammation. The same measurements can also help evaluate treatment-associated immune changes, provided the populations are defined consistently across samples.