Page 28 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

Conclusions

The IncuCyte Live-Cell Analysis System, in conjunction with • Proliferation assays can be run in microplates (96-well and
proprietary software tools and IncuCyte’s NucLight nuclear- 384-well) with high precision and reproducibility. In 384-
labeling reagents, provide a flexible assay platform for kinetic well plates, a mix and read assay is exemplified whereby full
measurements of proliferation. These assays achieve quantitative concentration-response curves of 16 standard anti-proliferative
and reproducible analysis of both adherent and non-adherent agents were compared. In a single IncuCyte instrument, 6 x
cells, in monoculture and importantly, co-culture, and without 384-well plates can be monitored providing >2000 wells of
removing cells from the physiologically-relevant environment of parallel data acquisition.
a tissue culture incubator. Image-based methodologies give the
user the ability to monitor morphological changes in parallel with • All data and time points can be verified by inspecting individual
quantification, the combination of which is a powerful and unique images and/or time-lapse movies. Cell morphology observations
tool for detecting pharmacological or genetic manipulations that provide additional validation and insight into mechanistic
alter cell viability or function. differences between treatments or conditions.

• Kinetic, label-free confluence or cell count measurements • Non-adherent cell proliferation and clustering can be visualized
over several cell divisions can be generated using phase image due to the non-perturbing nature of the IncuCyte’s optical
segmentation and without using fluorescent labels– an ideal design. Non-adherent cells stay stationary; there is no stage or
strategy for mono-culture analysis. sample movement, rather the optics move.

• Fluorescent reagents that label nuclei (e.g., IncuCyte NucLight
reagents) can be used to measure proliferation of mono-
cultures when cells are of low contrast and therefore difficult
to identify via segmentation of phase contrast images.
Nuclear-labeling strategies can also be used to differentiate the
effects of supporting cells on the proliferation of a labelled cell
population.

• IncuCyte’s kinetic label-free cell counting method can also
differentiate between proliferation rates of subpopulations
in co-cultures when individual cell types are of different size
or shape (non-adherent cells only), or when the individual
populations can be identified using cell surface markers
(adherent or non-adherent cells), or when one population is
fluorescently labelled with a nuclear marker (IncuCyte NucLight
reagents).

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