Description

The Cell Health Profiling BioApplication is an automated image analysis algorithm that provides detailed analysis of phenotypes related to cell health. This flexible tool allows you to analyze different features of interest, then create an activity profile based on patterns of features. These patterns, termed “Events”, can be defined to detect and distinguish, for example,

• Early and late cytotoxicity
• Apoptosis and necrosis
• Survival of target-expressing cells vs. non-expressors

In the example shown in Figure 1, cells were fluorescently labeled such that their nuclear morphology, mitochondrial potential (ΔΨµ), and plama membrane permeability could be assessed. The image analysis algorithm was configured to measure and report the presence of each featurespecific dye in the cells. A population of healthy cells was analyzed first to establish normal ranges for each feature, then thresholds were set to define Responder cells (cells with a value outside the range established by the control population).

Unique to this application is the ability to further classify the Responders based on the pattern of activity. An interactive wizard allows you to define
Events based on patterns of responses. In this example, the following Events were defined (see Figure 2):

• Event 1: Decrease in ΔΨµ AND NOT nuclear condensation AND NOT increased membrane permeability (identifies early toxicity)
• Event 2: Nuclear condensation OR increased membrane permeability (identifies moderate, irreversible toxicity)
• Event 3: Nuclear condensation AND increased membrane permeability (identifies late-stage, irreversible toxicity).

Features

• Enables definition of activity profiles or ‘Events’ at the level of individual cells
• Flexible configuration of cellular measurement regions (nucleus, organelles, whole cell)
• Performs measurements of up to 6 distinct targets
• Automatically characterizes patterns of activity at the single-cell and well-population levels
• Subpopulation definition and comparison
• Works with Cytotoxicity, Apoptosis, and Cell Viability HitKit™ HCS Reagent Kits

Benefits

• Enables distinction among cellular processes, for example apoptosis and necrosis
• Decreases time and effort required to determine toxic mechanisms of compounds
• Flexibility to choose cell health related targets and cell lines of interest
• Provides higher-quality information on cell subpopulations than conventional cell-based assays
• With HitKit reagent kits, provides a suite of turn-key cell health assays in one easy application

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