Acridine Orange Propidium Iodide
Acridine Orange Propidium Iodide - Related Videos
09:09
Annexin V and Propidium Iodide Labeling
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2023
Staining with annexin V and propidium iodide (PI) provides researchers with a way to identify different types of cell death—either necrosis or apoptosis. This technique relies on two components. The first, annexin V, is a protein that binds certain phospholipids called phosphatidylserines, which normally occur only in the inner, cytoplasm-facing leaflet of a cell’s membrane, but become “flipped” to the outer leaflet during the early stages of apoptosis. The second component is the DNA-binding...
07:34
Modified Annexin V/Propidium Iodide Apoptosis Assay For Accurate Assessment of Cell Death
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Cited by 543 •
2011
An accurate method for the assessment of cell death is described. The protocol improves upon conventional Annexin V/ propidium iodide (PI) protocols, which display up to 40% false- positive events in cell lines and primary cells from a broad range of animal models.
04:31
Differential Nuclear Staining Assay: An Assay to Determine Mitocan Cytotoxicity by Propidium Iodide and Hoechst Double Staining
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2025
This video demonstrates a nuclear staining method to detect the cytotoxic effect of mitocan on drug-sensitive cancer cells and healthy variants. The double staining with DNA-binding dyes such as propidium iodide and Hoechst dye provides a clear distinction between the dead and live cells, thus helping to assess the cytotoxicity of the drug.
08:53
Assessment of Neuronal Viability Using Fluorescein Diacetate-Propidium Iodide Double Staining in Cerebellar Granule Neuron Culture
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Cited by 29 •
2017
This protocol describes how to accurately measure neuronal viability using Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining in cultured cerebellar granule neurons, a primary neuronal culture used as an in vitro model in neuroscience and neuropharmacology research.
04:18
DNA Electrophoresis Using Thiazole Orange Instead of Ethidium Bromide or Alternative Dyes
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Cited by 5 •
2019
Here, we present a protocol to use thiazole orange for the detection of DNA in gel electrophoresis experiments. The use of thiazole orange allows elimination of ethidium bromide, and fluorescence detection can be achieved with either UV or blue light.
FAQs
Acridine orange marks nucleic acid-containing cells green because it can cross both intact and damaged plasma membranes. Propidium iodide produces red fluorescence only when membrane damage permits entry, after which it binds DNA or RNA. Comparing the two signals therefore links fluorescence patterns to membrane status rather than treating either dye as a standalone indicator.
The decisive information comes from the relationship between signals: green fluorescence indicates nucleic-acid staining by acridine orange, whereas red fluorescence indicates that propidium iodide reached nucleic acids after membrane compromise. Examining both channels helps separate cells with preserved membranes from those with damage and makes treatment-associated shifts in population health easier to interpret.
A visual readout allows researchers to inspect fluorescence patterns directly, whereas a flow-based readout supports analysis of the same stained population through fluorescence signals. Both approaches use the complementary behavior of the dyes, but they serve different reporting needs: visual inspection emphasizes rapid assessment, while flow analysis supports population-level evaluation of cell health and treatment effects.
Membrane integrity is important because it determines whether propidium iodide can enter the cell. In apoptosis-related analyses, the red signal can therefore contribute to evaluating changes associated with cell damage, while acridine orange supplies the complementary nucleic-acid signal. The assay is consequently useful for relating treatment effects to altered cell viability patterns.
For cytotoxicity experiments, researchers can compare fluorescence patterns in cell populations exposed to different treatments. A greater representation of propidium iodide-associated red fluorescence indicates more cells with compromised membranes, while the paired green signal helps assess the remaining stained population. This comparison provides a rapid readout of treatment-associated changes in cell health.
In infection studies, the assay can be used to evaluate pathogen-induced cellular damage by examining shifts in the green and red fluorescence pattern. Because the readout reflects membrane status across a population, it helps connect infection conditions with changes in cell health. This makes the method relevant for assessing how strongly a pathogen-associated process affects host cells.
Dual fluorescence supports cell counting while also indicating the health distribution of the measured population. Researchers can use the contrasting signals to distinguish cells with preserved membranes from cells showing membrane-associated damage, then assess how that distribution changes after treatment or infection. The resulting pattern provides a rapid population-level view of experimental outcomes.