Annexin V Propidium Iodide
Annexin V 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.
04:21
Annexin V Binding Assay: A Fluorescence-Based Technique to Identify Apoptotic Erythrocytes via Phosphatidylserine Labeling
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2025
In this video, we demonstrate the Annexin V binding assay to measure the degree of eryptosis, erythrocyte programmed cell death, using fluorescent Annexin V staining. Apoptotic erythrocytes demonstrate phosphatidylserine translocation from the inner to the outer leaflet of the cell membrane, causing the fluorophore-tagged Annexin V to bind to the translocated phosphatidylserine, thereby emitting a fluorescent signal.
FAQs
The two signals report different membrane events rather than duplicating one another. Annexin V positivity indicates that phosphatidylserine has become exposed externally, an early-apoptotic change. Propidium iodide positivity indicates loss of plasma-membrane integrity. Reading both channels together therefore distinguishes cells with early apoptotic changes from those that have progressed to late apoptosis or necrosis.
Interpretation depends on the combination of signals. Annexin V-negative and propidium iodide-negative cells are classified as viable, whereas Annexin V-positive and propidium iodide-negative cells represent early apoptosis. Cells positive for both markers indicate late apoptosis, while propidium iodide-positive cells without Annexin V signal are classified as necrotic. This enables quantitative comparison of cell-death states.
A phosphatidylserine signal alone identifies an apoptotic membrane change but does not indicate whether the plasma membrane remains intact. Propidium iodide adds that information by marking membrane compromise. The combined readout therefore provides greater discrimination between early apoptotic cells, cells with advanced death-associated membrane damage, and necrotic populations than either signal considered independently.
After staining, investigators detect the two fluorescence signals using either flow cytometry or fluorescence microscopy. Flow cytometry is especially useful for separating and quantifying viable, apoptotic, and necrotic populations within a sample. Microscopy provides a fluorescence-based way to examine staining at the cellular level, supporting direct assessment of cell-death profiles.
The workflow involves applying Annexin V and propidium iodide to cells, detecting their distinct fluorescence signals, and assigning cells to viability or death categories according to the combined profile. Researchers can then compare the relative sizes of viable, early-apoptotic, late-apoptotic, and necrotic populations. This produces a rapid quantitative assessment of cell viability and death.
The assay is useful when researchers need to measure how biological or experimental conditions alter cell survival and death. Applications include studying development, investigating disease mechanisms, evaluating cytotoxicity, and assessing drug responses. Because it separates several cell-death states, the method can show whether a treatment primarily preserves viability or shifts cells toward apoptosis or necrosis.