Source 1review1992Cytometry
Toolkit/Acridine orange supravital lysosomal proton pump assay
Acridine orange supravital lysosomal proton pump assay
Also known as: acridine orange uptake assay
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The ATP-dependent lysosomal proton pump, tested by the supravital uptake of acridine orange (AO) was also preserved in apoptotic but not necrotic cells.
Usefulness & Problems
Why this is useful
This assay uses supravital acridine orange uptake to test whether the ATP-dependent lysosomal proton pump is preserved.; testing lysosomal proton pump preservation; distinguishing apoptotic from necrotic cells
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This assay uses supravital acridine orange uptake to test whether the ATP-dependent lysosomal proton pump is preserved.
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testing lysosomal proton pump preservation
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distinguishing apoptotic from necrotic cells
Problem solved
It adds a lysosomal-function dimension for distinguishing apoptotic from necrotic cells.; provides a lysosomal function readout that differs between apoptotic and necrotic cells
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It adds a lysosomal-function dimension for distinguishing apoptotic from necrotic cells.
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provides a lysosomal function readout that differs between apoptotic and necrotic cells
Problem links
provides a lysosomal function readout that differs between apoptotic and necrotic cells
LiteratureIt adds a lysosomal-function dimension for distinguishing apoptotic from necrotic cells.
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It adds a lysosomal-function dimension for distinguishing apoptotic from necrotic 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
atp-dependent lysosomal proton pumpinglysosomal acidification-dependent dye accumulationsupravital dye uptakeTechniques
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 acridine orange and flow-cytometric fluorescence measurement.; requires acridine orange supravital uptake; depends on flow-cytometric fluorescence measurement
The abstract does not state that it independently stages apoptosis or identifies DNA damage features.
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-supravital-lysosomal-proton-pump-assay
The ATP-dependent lysosomal proton pump, tested by the supravital uptake of acridine orange (AO) was also preserved in apoptotic but not necrotic cells.
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review summarysupports
Supravital acridine orange uptake indicates that ATP-dependent lysosomal proton pump function is preserved in apoptotic but not necrotic cells.
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Comparisons
Source-stated alternatives
Other methods in the review include PI/Hoechst staining, rhodamine 123 retention, DNA content analysis, low-pH acridine orange denaturation, and nick translation.
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Other methods in the review include PI/Hoechst staining, rhodamine 123 retention, DNA content analysis, low-pH acridine orange denaturation, and nick translation.
Source-backed strengths
reports preservation of ATP-dependent lysosomal proton pump activity in apoptotic but not necrotic cells
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reports preservation of ATP-dependent lysosomal proton pump activity in apoptotic but not necrotic cells
Compared with Langendorff perfused heart electrical recordings
Acridine orange supravital lysosomal proton pump assay and Langendorff perfused heart electrical recordings address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Compared with native green gel system
Acridine orange supravital lysosomal proton pump assay and native green gel system address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Acridine orange supravital lysosomal proton pump assay and sub-picosecond pump-probe analysis of bacteriorhodopsin pigments address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.