Page 15 - Live-cellanalysis handbook

Kinetic Cell Health and Viability Assays

Real-time automated measurements
of cell health and viability

Measurements of cell health are essential for studying the effects How Live-Cell Health and Viability Assays Work
of drugs, culture conditions or genetic modifications on cell
growth or viability. Such studies are used to rank compounds in Proliferation and viability assays
drug discovery screens, identifying off-target toxic compounds, as
well as to investigate the cellular changes that underline disease Measure growth or growth inhibition in real time over several cell
pathologies. In order to assess cell health and viability, a variety divisions using label-free cell counts or confluence measurements
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of end-point assays have been employed, such as ATP assays, in both adherent and non-adherent cell types. Additional assay
LDH assays and vital dyes used in flow cytometry, but all fail to strategies can be used to generate measurements of cell type-
take into account the kinetics or have the capability to make specific growth rates in co-cultures. IncuCyte NucLight reagents
®
simultaneous measurements in a single well. Understanding the can be used to fluorescently label nuclei and determine counts of
biological time-dependence, along with the ability to evaluate viable cells.
multiple methods by which a cell dies, offers considerable
advantages in the characterization of test compounds. Apoptosis assays
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IncuCyte® cell health and viability assays allow for the kinetic Detect apoptosis in living cells and in real time using mix-and-read
evaluation of effective concentration of the compound and the reagents that measure caspase-3/7 activity or phosphatidyl serine
time needed for the compound to perturb the target, thereby externalization.
discriminating if the test agent is fast acting or prolonged. Another
advantage is the ability to perform multi-parametric analysis Cytotoxicity assays
using non-perturbing reagents within a single well, enabling
detection of the mechanism of cell death as well as monitoring cell Measure cell death upon loss of plasma membrane integrity over
viability utilizing reagents that label cell nuclei. These image-based time using simple mix-and-read protocols.
detection methods can all be qualitatively validated with images to
evaluate morphological changes associated with cell death.

Furthermore, these advantages of live-cell analysis are also ideally
suited to meet the needs of more dynamic cellular models, such
as co-culture immune cell killing assays, which require robust
temporal and spatial quantitative measurements in physiologically-
relevant conditions. Continuous live-cell imaging makes it possible
to measure and visually validate the complex and dynamic
interactions between target and effector cells in co-culture for the
reliable analysis of new potential immunotherapies.

Live-cell imaging and analysis approaches

The IncuCyte cell health and viability assays enable the detection
and analysis of cell proliferation, viability, apoptosis, and
cytotoxicity. Images of cell cultures in 96- or 384-well formats
are automatically acquired and analyzed to generate time courses
and reveal concentration-dependent responses that can be used to
calculate EC50 or IC50 values.

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