Lymphocyte Gating
Lymphocyte Gating - Related Videos
03:23
An In Vitro Technique to Stimulate Lymphocytes via Pathogenic Bacteria
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2025
This video demonstrates an in vitro technique to stimulate lymphocytes via non-typeable Haemophilus influenzae (NTHi) and to assess the immune response. Incubating isolated peripheral blood mononuclear cells (PBMCs) with NTHi causes the bacterial cells to stimulate T lymphocytes, leading to cytokine production. Upon inhibiting the secretion of cytokines, immunofluorescence staining is performed to detect the accumulated intracellular cytokines in stimulated T lymphocytes.
04:46
Isolation of Lymphocytes from Mouse Genital Tract Mucosa
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Cited by 10 •
2012
An efficient way to isolate lymphocytes from mouse genital tract is described. This method takes advantage of enzyme digestion and Percoll gradient separation to allow efficient isolation. This technique is also adaptable to for use in other...
04:45
An Immunological Technique to Monitor Adjuvant-Mediated Cytotoxic T Lymphocyte Generation
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2025
The video demonstrates an immunological method to monitor in vivo adjuvant-induced cytotoxic T lymphocyte or CTL production. A mouse is pre-injected with genetically engineered, proliferative dye-stained donor CD8+ T lymphocytes. Subcutaneous injection of ovalbumin with adjuvants generates cytotoxic CD8+ T lymphocytes that are identified post-harvesting through immunostaining and flow cytometry.
12:55
P50 Sensory Gating in Infants
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Cited by 5 •
2013
Methods to record auditory P50 sensory gating, a physiological marker of cerebral inhibition which reflects early stages of attention, are described.
12:59
Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes
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Cited by 33 •
2013
With effector functions distinct from other T cell subsets, Th17 cells have been centrally implicated in inflammatory autoimmunity. This in vitro Th17 differentiation protocol provides a means to determine whether naïve CD4+ T lymphocytes can differentiate into Th17 cells, and to further examine their role in autoimmunity and host response.
FAQs
Sequential gating narrows analysis in stages: first, cells are selected using physical scatter characteristics, then fluorescence signals refine the population by marker expression. This order helps remove debris and unrelated leukocyte populations before interpreting T-cell, B-cell, or natural killer cell measurements. It therefore improves confidence that frequencies and phenotypic differences reflect the intended lymphocyte group rather than the mixed sample.
Scatter signals help identify cells with lymphocyte-like size and granularity, providing an initial separation from other material in the sample. Fluorescence adds biological specificity by detecting markers such as CD3, CD19, or CD56. Using both types of information is important because physical appearance alone does not distinguish the major lymphocyte lineages needed for focused immune analysis.
These surface markers provide complementary information about lymphocyte identity and phenotype. CD3 supports identification of T cells, while CD19 identifies B cells and CD56 identifies natural killer cells; CD4 and CD8 further characterize T-cell populations. Examining these signals allows researchers to measure the composition of the selected lymphocyte population rather than treating it as a single uniform group.
Debris, dead cells, and other leukocyte populations can enter the analyzed region if gates are not carefully chosen. Their inclusion may alter calculated immune-cell frequencies and obscure genuine differences in phenotype or activation state. Excluding these unwanted events through sequential selection makes downstream comparisons more reliable, particularly when samples contain diverse cellular material during immunology or infection studies.
A typical workflow begins with a mixed cell sample and uses forward- and side-scatter properties to select events with lymphocyte-like size and granularity. Fluorescence signals are then applied to distinguish populations through markers such as CD3, CD4, CD8, CD19, and CD56. Researchers can subsequently assess selected cells for frequency, phenotype, activation state, or responses to pathogens or treatment.
In immunology and infection studies, the strategy focuses measurements on relevant immune-cell populations within a mixed sample. Researchers can quantify T cells, B cells, and natural killer cells, then examine changes in their phenotypes, activation states, or responses to pathogens and treatment. This focused analysis helps connect cellular patterns with immune responses without allowing unrelated events to dominate the results.
Accurate gating supports comparisons of immune-cell frequencies and marker-defined phenotypes across samples or experimental conditions. It can also reveal differences in activation states and responses associated with pathogens or treatment. Because the analysis excludes debris, dead cells, and other leukocytes, observed changes are more likely to represent the selected lymphocyte populations and to support dependable downstream conclusions.