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Real-Time, Automated Cytotoxicity Assays within a Tissue Culture Incubator

Tuesday, February 6, 2018   (0 Comments)
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Cytotoxicity
The detrimental effects caused by substances or environmental changes on cell health are described as cytotoxicity. When cells are exposed to a cytotoxic stimulus, their metabolic activity may be compromised and cell growth or division may be inhibited, eventually leading to cell death.

Apoptotic cell death involves a more controlled and programmed mechanism, whereas necrotic cell death is a catastrophic cell lysis. A specialized process involving digestion of cells by themselves from within is called autophagy. Cell death is inevitable when there is an irreversible loss of membrane integrity for a cell.

Cytotoxicity assays
In most cytotoxicity assays, cell membrane integrity is measured either by markers released from dying cells (for instance, cellular proteases) or with vital dyes that are removed from healthy cells, for instance, propidium iodide or trypan blue.

Cell viability and health is measured using metabolic activity measurements, for instance ATP, LDH, or MTT assays. However, only a few methods are available that can directly count the number of dying cells over time.

IncuCyte® cytotoxicity assay
Real-time, automated cytotoxicity assays can be achieved within a tissue culture incubator using the IncuCyte® s3 live-cell imaging and analysis system. Using the IncuCyte® Cytotox reagents, cell death is measured in real-time based on cell membrane integrity by the IncuCyte™ Cytotoxicity Assay, all within a tissue culture incubator.

IncuCyte® Cytotox Red Reagent - labels dying cells red
IncuCyte® Cytotox Green Reagent - labels dying cells green
The IncuCyte® Cytotox reagents are non-fluorescent, inert, and do not enter healthy cells when introduced into the tissue culture growth medium. However, the Cytotox probe enters death cells due to the loss of membrane integrity, fluorescently labeling the nuclei.

Based on the appearance of red (or green) labeled nuclei, the probe allows identification and quantification of dying cells over time. Cell death can be further validated based on morphology using high definition phase contrast movies and images (for example, loss of motility and loss of cytoskeleton structure).

 

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