Cytotoxicity Assays for Live-Cell Analysis

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What is Cytotoxicity?

Cytotoxicity is a general term that describes the detrimental effects of substances or environmental changes on cell health. Exposure of cells to a cytotoxic stimulus may compromise metabolic activity, inhibit cell growth or division or ultimately produce cell death. 'Necrotic' cell death is a catastrophic cell lysis. 'Apoptotic' cell death is a more controlled, programmed mechanism. 'Autophagy' is a specialised process whereby cells digest themselves from within. Irrespective of the mechanism of cytotoxicity, once a cell irreversibly loses its membrane integrity it is destined to die.

A number of cytotoxicity assays involve measurement of cell membrane integrity, either with vital dyes that are excluded from healthy cells, e.g. trypan blue or propidium iodide, or via the release of markers from dying cells (e.g. cellular proteases). Metabolic activity measurements, e.g. MTT, LDH or ATP assays are also used to measure cell health and viability. However, despite these methods, few if any involve direct counting of the number of dying cells over time.
 

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Introducing the Incucyte® Cytotoxicity Assay

The Incucyte® Live-Cell Analysis System enables real-time, automated cytotoxicity assays within your tissue culture incubator.

The Incucyte® Cytotoxicity Assay uses the Incucyte® Cytotox Dyes to make real time measurements of cell death based on cell membrane integrity, all within your incubator.

  • Incucyte® Cytotox Green Dye*** - labels dying cells green
  • Incucyte® Cytotox Red Dye*** - labels dying cells red

When added to the tissue culture growth medium, the Incucyte® Cytotox Dyes are inert, non-fluorescent and do not enter viable cells. As cells die and membrane integrity is lost, the Cytotox Dye enters the cell and fluorescently labels the nuclei. Dying cells are identified and quantified over time by the appearance of green- (or red) labelled nuclei. High definition phase contrast images and movies provide an additional validation of cell death based on morphology (e.g. loss of cytoskeleton structure, loss of motility).

Measure cytotoxicity in tumor, immune or neuronal cultures using the Incucyte® Cytotoxicity Assay. (Left) Real-time detection of cytotoxicity in HT-1080 fibrosarcoma cells following treatment with the cytotoxic drug camptothecin. Dying cells are labeled green, in real-time, by the mix-and-read Incucyte® Cytotox Green Dye. (Right) Incucyte® Cytotox Red Dye labels dead rat forebrain neurons treated with 333 μM glutamate.

Incucyte® Cytotoxicity Assay Concept

  1. Detect cytotoxicity in real-time by adding the mix-and-read Incucyte® Cytotox Dyes to your cultures
  2. Automatically quantify number of dying cells using intuitive Incucyte® live-cell imaging and analysis tools
  3. Multiplex cytotoxicity readouts with measurements of proliferation or apoptosis by combining with Incucyte® Nuclight nuclear labeling reagents, the Incucyte® Caspase 3/7 Dye, or the Incucyte® Annexin V Dye

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