Source 1review1992Cytometry
Toolkit/Acridine orange low-pH DNA denaturation assay
Acridine orange low-pH DNA denaturation assay
Also known as: acridine orange DNA denaturation assay, AO low-pH DNA denaturation assay
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The sensitivity of DNA in situ to denaturation, was increased in apoptotic and necrotic cells. This feature, probed by staining with AO at low pH, provided a sensitive and early assay to discriminate between live, apoptotic and necrotic cells, and to evaluate the cell cycle phase specificity of these processes.
Usefulness & Problems
Why this is useful
This assay probes increased in situ DNA denaturation sensitivity using acridine orange at low pH to separate live, apoptotic, and necrotic cells.; early discrimination of live, apoptotic, and necrotic cells; evaluating cell cycle phase specificity of cell death processes
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This assay probes increased in situ DNA denaturation sensitivity using acridine orange at low pH to separate live, apoptotic, and necrotic cells.
Source:
early discrimination of live, apoptotic, and necrotic cells
Source:
evaluating cell cycle phase specificity of cell death processes
Problem solved
It offers an early and sensitive way to classify cell death state while also supporting analysis of cell-cycle phase specificity.; provides a sensitive early assay for distinguishing cell death states and relating them to cell cycle phase
Source:
It offers an early and sensitive way to classify cell death state while also supporting analysis of cell-cycle phase specificity.
Source:
provides a sensitive early assay for distinguishing cell death states and relating them to cell cycle phase
Problem links
provides a sensitive early assay for distinguishing cell death states and relating them to cell cycle phase
LiteratureIt offers an early and sensitive way to classify cell death state while also supporting analysis of cell-cycle phase specificity.
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It offers an early and sensitive way to classify cell death state while also supporting analysis of cell-cycle phase specificity.
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
acid-induced dna denaturation sensitivity readoutdifferential acridine orange staining of in situ dna under low-ph conditionsTechniques
Functional AssayTarget processes
recombinationImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
It requires acridine orange, low-pH staining conditions, and flow-cytometric readout.; requires acridine orange staining at low pH; depends on flow-cytometric analysis
The abstract does not say that it directly measures membrane integrity or mitochondrial potential.
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 acridine-orange-low-ph-dna-denaturation-assay
The sensitivity of DNA in situ to denaturation, was increased in apoptotic and necrotic cells. This feature, probed by staining with AO at low pH, provided a sensitive and early assay to discriminate between live, apoptotic and necrotic cells, and to evaluate the cell cycle phase specificity of these processes.
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review summarysupports
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.
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Comparisons
Source-stated alternatives
The review also presents PI/Hoechst staining, rhodamine 123 retention, acridine orange uptake for lysosomal function, DNA content measurements, and in situ nick translation.
Source:
The review also presents PI/Hoechst staining, rhodamine 123 retention, acridine orange uptake for lysosomal function, DNA content measurements, and in situ nick translation.
Source-backed strengths
described as sensitive and early
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described as sensitive and early
Compared with barcoded Cre recombinase mRNA barcode platform
Acridine orange low-pH DNA denaturation assay and barcoded Cre recombinase mRNA barcode platform address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination
Compared with calcium imaging
Acridine orange low-pH DNA denaturation assay and calcium imaging address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination
Relative tradeoffs: appears more independently replicated.
Compared with two-photon excitation microscopy
Acridine orange low-pH DNA denaturation assay and two-photon excitation microscopy address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.