Source 1review1992Cytometry
Toolkit/Propidium iodide and Hoechst 33342 apoptosis discrimination assay
Propidium iodide and Hoechst 33342 apoptosis discrimination assay
Also known as: PI/Hoechst 33342 assay, propidium iodide followed by Hoechst 33342
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The combination of PI followed by Hoechst 33342 proved to be an excellent probe to distinguish live, necrotic, early- and late-apoptotic cells.
Usefulness & Problems
Why this is useful
This assay combines propidium iodide exclusion with Hoechst 33342 staining to separate live, necrotic, early-apoptotic, and late-apoptotic cells by flow cytometry.; distinguishing live, necrotic, early-apoptotic, and late-apoptotic cells; probing plasma membrane integrity by flow cytometry
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This assay combines propidium iodide exclusion with Hoechst 33342 staining to separate live, necrotic, early-apoptotic, and late-apoptotic cells by flow cytometry.
Source:
distinguishing live, necrotic, early-apoptotic, and late-apoptotic cells
Source:
probing plasma membrane integrity by flow cytometry
Problem solved
It addresses the need to distinguish apoptosis from necrosis and to stage apoptotic progression rather than only scoring dead versus live cells.; discriminates apoptosis from necrosis using membrane integrity and nuclear staining patterns
Source:
It addresses the need to distinguish apoptosis from necrosis and to stage apoptotic progression rather than only scoring dead versus live cells.
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discriminates apoptosis from necrosis using membrane integrity and nuclear staining patterns
Problem links
discriminates apoptosis from necrosis using membrane integrity and nuclear staining patterns
LiteratureIt addresses the need to distinguish apoptosis from necrosis and to stage apoptotic progression rather than only scoring dead versus live cells.
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It addresses the need to distinguish apoptosis from necrosis and to stage apoptotic progression rather than only scoring dead versus live cells.
Published Workflows
Objective: Differentiate apoptosis from necrosis and identify early apoptotic changes and cell-cycle phase specificity using multiparameter flow-cytometric readouts.
Why it works: The review describes combining orthogonal cellular features so that apoptosis and necrosis can be separated by differences in DNA behavior, membrane integrity, organelle function, and scatter properties rather than relying on a single marker.
endonuclease-associated low molecular weight DNA extractionloss or preservation of plasma membrane integritypreservation or loss of mitochondrial transmembrane potentialpreservation or loss of ATP-dependent lysosomal proton pumpingDNA strand-break formationflow cytometrymultiparameter fluorescent stainingbivariate analysis
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
differential dye exclusion/permeability stainingfluorescence-based cell-state discriminationTechniques
Functional AssayTarget processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
It requires PI, Hoechst 33342, and a flow cytometer capable of measuring the corresponding fluorescence signals.; requires propidium iodide and Hoechst 33342 staining; depends on flow-cytometric measurement
The abstract does not state that it identifies the molecular trigger of death or replaces other orthogonal apoptosis measurements.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Acridine orange staining at low pH provides a sensitive and early assay to discriminate live, apoptotic, and necrotic cells and to evaluate cell-cycle phase specificity.
Propidium iodide exclusion combined with Hoechst 33342 staining is described as an excellent probe for distinguishing live, necrotic, early-apoptotic, and late-apoptotic cells.
Rhodamine 123 retention indicates that mitochondrial transmembrane potential is preserved in apoptotic but not necrotic cells.
Supravital acridine orange uptake indicates that ATP-dependent lysosomal proton pump function is preserved in apoptotic but not necrotic cells.
Approval Evidence
1 source1 linked approval claimfirst-pass slug propidium-iodide-and-hoechst-33342-apoptosis-discrimination-assay
The combination of PI followed by Hoechst 33342 proved to be an excellent probe to distinguish live, necrotic, early- and late-apoptotic cells.
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review summarysupports
Propidium iodide exclusion combined with Hoechst 33342 staining is described as an excellent probe for distinguishing live, necrotic, early-apoptotic, and late-apoptotic cells.
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Comparisons
Source-stated alternatives
The review also discusses DNA content measurements, rhodamine 123 retention, acridine orange uptake or low-pH denaturation assays, scatter changes, and in situ nick translation.
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The review also discusses DNA content measurements, rhodamine 123 retention, acridine orange uptake or low-pH denaturation assays, scatter changes, and in situ nick translation.
Source-backed strengths
described as an excellent probe for multi-state cell death discrimination
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described as an excellent probe for multi-state cell death discrimination
Compared with assays
The review also discusses DNA content measurements, rhodamine 123 retention, acridine orange uptake or low-pH denaturation assays, scatter changes, and in situ nick translation.
Shared frame: source-stated alternative in extracted literature
Strengths here: described as an excellent probe for multi-state cell death discrimination.
Source:
The review also discusses DNA content measurements, rhodamine 123 retention, acridine orange uptake or low-pH denaturation assays, scatter changes, and in situ nick translation.
Ranked Citations
- 1.