Propidium Iodide Staining
Propidium Iodide Staining - 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...
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.
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.
04:38
Assessing Neuronal Viability in a Cerebellar Neuron Culture via Double Staining
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2025
This video demonstrates the method of identifying neurons from a mixed culture of cerebellar granule neurons and glial cells using dual staining with fluorescein diacetate (FDA) and propidium iodide (PI), where live neurons appear green and dead neurons appear red.
FAQs
Signal intensity is governed primarily by membrane access, not simply by the presence of nucleic acids. Intact cells exclude propidium iodide, whereas membrane-compromised cells permit entry and retain the dye through nucleic-acid intercalation. This makes the red-fluorescent population a readout of membrane disruption, which is especially relevant when evaluating neuronal damage.
Microscopy and flow cytometry answer related but different measurement needs. Microscopy shows where propidium iodide-positive cells occur and can connect fluorescence with cellular or tissue morphology, while flow cytometry measures fluorescence across many cells for population-level quantification. Choosing between them depends on whether spatial context or numerical cell-population analysis is the main objective.
Annexin V adds information about cell-state interpretation that propidium iodide alone cannot provide. In a combined assay, cells retaining membrane exclusion can be compared with cells that also become propidium iodide positive. This pattern helps separate viable cells from membrane disruption associated with late apoptosis or necrosis, improving interpretation of neuronal cell-death measurements.
A positive signal should be interpreted as evidence of compromised membrane integrity rather than as a standalone label for one specific death pathway. Membrane disruption can accompany late apoptosis or necrosis, so the fluorescence identifies an affected-cell population but does not by itself distinguish those outcomes. Pairing it with Annexin V provides the additional context.
A basic workflow consists of applying propidium iodide to the cells or tissue sample, detecting red fluorescence with microscopy or flow cytometry, and quantifying the fluorescent population. The selected readout should match the question: imaging supports location and visual assessment, whereas flow cytometry supports numerical analysis of cell populations.
In neuronal cultures, the assay can track loss of membrane integrity after an injury or treatment that may produce neurotoxic effects. In tissue samples, fluorescence measurements can identify and quantify affected cells within the preparation. These applications make the method useful for comparing neuronal damage or cell loss across experimental conditions.