Pyroptosis Detection
Pyroptosis Detection - Related Videos
05:14
Examination of Pyroptosis by Flow Cytometry
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2024
This article describes the identification of pyroptotic cells using flow cytometry after dual staining with antibodies against the N-terminal fragment of chicken GSDME (chGSDME-NT) and propidium iodide (PI).
03:02
A Lactate Dehydrogenase Release Assay To Detect Macrophage Death Following Nigericin Exposure
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2025
This video demonstrates the lactate dehydrogenase assay to detect inflammasome-mediated macrophage death. The macrophages are exposed to lipopolysaccharide for priming, followed by exposure to nigericin to cause inflammasome-induced cell death, which is determined by the lactate dehydrogenase assay.
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.
09:10
Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce
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Cited by 17 •
2010
This protocol describes a simple adhesive-tape-based approach for sampling of tomato and other fresh produce surfaces, followed by rapid whole cell detection of Salmonella using fluorescence in situ hybridization (FISH).
09:00
Detection of Protein Ubiquitination
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Cited by 96 •
2009
Ubiquitination is a key posttranslational modification carried out by a set of three enzymes. Mutations of genes involved in this modification are associated with many different human diseases. Here, we describe protocols to detect protein ubiquitination in cultured cells in vivo and test tubes in vitro.
FAQs
Reliable interpretation depends on combining upstream and downstream evidence rather than treating one signal as conclusive. Researchers can examine inflammatory caspase activation, gasdermin D cleavage and pore formation, then compare those findings with membrane permeabilization, propidium iodide uptake, lactate dehydrogenase release, or cytokine secretion. Concordant results strengthen attribution to pyroptosis.
Inflammasomes provide a mechanistic link between infection or cellular danger signals and inflammatory cell death. Their activation is evaluated through evidence of inflammatory caspase activity, followed by gasdermin D cleavage and pore formation. Measuring these events alongside interleukin-1β or interleukin-18 secretion helps connect intracellular sensing with downstream inflammatory output.
Distinction requires evaluating a coordinated pattern rather than relying only on general membrane damage. Inflammatory caspase activation, gasdermin D cleavage, pore formation, and inflammatory cytokine release provide pathway-related evidence, while propidium iodide uptake or lactate dehydrogenase release indicate membrane disruption. Comparing these readouts helps determine whether inflammation accompanies the observed cell death.
Propidium iodide uptake and lactate dehydrogenase release are downstream indicators of membrane permeabilization. They can be interpreted with evidence of gasdermin D pore formation to connect loss of membrane integrity with the pyroptotic mechanism. Interleukin-1β and interleukin-18 secretion adds an inflammatory dimension, helping assess whether membrane damage is associated with immune signaling.
A practical workflow begins by assessing inflammatory caspase activation and gasdermin D cleavage, followed by evaluation of pore formation or membrane permeabilization. Researchers then measure propidium iodide uptake, lactate dehydrogenase release, or secretion of interleukin-1β and interleukin-18. Using several stages produces a more informative profile than measuring a single endpoint.
The approach is useful when researchers need to determine how pathogens or cellular danger signals activate inflammasomes and inflammatory cell death. It can reveal whether infection is associated with membrane disruption and cytokine release, helping characterize host responses and potential inflammatory damage. These measurements also support comparisons between different infection-related conditions.
Pyroptosis-related readouts can show whether a treatment changes inflammatory caspase activation, gasdermin D cleavage, pore formation, membrane permeabilization, or cytokine secretion. Measuring several endpoints indicates whether an intervention affects the pathway itself, its cellular consequences, or inflammatory output. This supports assessment of treatments intended to modulate host defense or inflammatory damage.