Inflammasome Visualization
Inflammasome Visualization - Related Videos
03:44
Detection of Inflammasome Activation via Fluorescence Microscopy
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2025
This video demonstrates a method to detect NLRP3 inflammasome activation in macrophages using fluorescent reporter probes. The cells are treated with potassium ionophores and glycine to induce inflammasome assembly and caspase-1 activation. Fluorescent reporter probes visualize activated caspase-1, and cells are stained with a fluorescent dye for nuclei visualization.
03:06
An In Vitro Technique to Study Inflammasome Complex Formation in Macrophages
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2025
This video demonstrates a technique to study NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome formation in primary macrophages via immunofluorescence. The macrophages are first primed with bacterial lipopolysaccharide (LPS) to upregulate NLRP3 protein expression. Exposing the macrophages to nigericin—a potassium ionophore—leads to the activation of NLRP3 and the formation of inflammasomes, which are visualized via immunofluorescence.
09:04
Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
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Cited by 7 •
2014
Dendritic cells (DCs) secrete IL-1β in response to TLR8 recognition of synthetic purine, R848, followed by NLRP3 inflammasome activation with nigericin, therefore, IL-1β can be used to measure NLRP3 inflammasome activity. Intracellular cytokine staining, immunoblotting, and ELISA are used to accurately measure NLRP3 inflammasome priming and activation via IL-1β expression.
05:19
Visualization of Caspase Activation in Human Monocyte-Derived Macrophages
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2025
This video demonstrates the activation of caspase proteins in human macrophages using bimolecular fluorescence complementation reporter proteins. The pro-domains of caspase-1 are fused with non-fluorescent fragments of Venus proteins, which are recruited to inflammasome complexes. The proximity of inflammasomes induces the refolding and fluorescence of Venus fragments, confirming caspase activation in the macrophages.
09:44
Methods to Test Visual Attention Online
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Cited by 26 •
2015
To replicate laboratory settings, online data collection methods for visual tasks require tight control over stimulus presentation. We outline methods for the use of a web application to collect performance data on two tests of visual attention.
FAQs
Activation can be followed as a sequence rather than treated as a single endpoint. After infection or cellular stress engages a sensor, ASC is recruited into a punctate complex, and inflammatory caspase-1 becomes associated with the resulting signaling state. Imaging therefore helps place complex formation before downstream cytokine maturation and pyroptotic cell death in time.
ASC puncta provide a visible readout of adaptor recruitment and complex organization. Their presence can indicate that a sensor has engaged the inflammasome pathway, while imaging their location and timing helps relate assembly to later caspase-1-associated activity. This makes ASC especially useful for connecting molecular organization with cell-level responses during infection or stress.
Because imaging records defined fluorescent events in individual cells, it can distinguish cells showing inflammasome-associated assembly from cells that display inflammation without that visual signature. This comparison is important when bulk measurements could combine responding and nonresponding populations. The approach therefore adds pathway-specific spatial information to immunology studies of infection and cellular stress.
Researchers can track where fluorescent signals appear and when they change, rather than relying only on a final measurement. Spatial information shows which cells or cellular locations contain relevant complexes, while time-resolved observation relates assembly to caspase-1 activation and subsequent outcomes. Together, these dimensions reveal how signaling unfolds across individual cells.
Imaging can be used to compare cells within the same sample and identify which populations exhibit inflammasome-associated fluorescent events. This is valuable when infection or stress affects only a subset of cells. Linking those visual responses with cytokine maturation or pyroptotic cell death helps researchers evaluate heterogeneity in host defense rather than interpreting the sample as uniformly activated.
In infection and disease-model studies, the method can reveal whether a pathogen changes host inflammasome responses or whether a candidate therapeutic alters them. Researchers can compare visualized assembly and activity with downstream cellular outcomes, including cytokine maturation and pyroptotic cell death. This supports evaluation of how interventions affect innate immune signaling and host defense.