Page 119 - Live-cellanalysis handbook

Kinetic Neuronal Activity Assays

Conclusions

The IncuCyte Neuronal Activity Assay, consisting of • Acquisition of longer wavelength fluorophores (excitation >
®
instrumentation, software, and reagent, provides access to 500 nm) allows for chronic analysis of the same population
complex, neuronal activity and connectivity measurements as of cells over weeks to months to detect functional changes
neurons and their networks mature and become functionally and reveal differences among various iPSC-derived neuronal
active through chronic evaluation of the same population of cells. models or to measure impact of culture conditions on
This assay achieves long-term evaluation in a variety of neuronal neuronal activity.
cell models, such as primary cells and hiPSC-derived neurons, to
evaluate when they become active and how this activity changes • Capture of spontaneous Ca2+ oscillations from thousands
overtime without removing cells from the physiologically-relevant of neurons allows for highly reproducible kinetic profiling
environment of a tissue culture incubator. Overall, live-cell analysis to determine both activity and connectivity using IncuCyte
provides a valuable compliment to established techniques such Neuronal Activity Analysis software tools.
as electrophysiology, for building and validating translational cell
models for the discovery of novel neurotherapeutics. • Additional insights can be obtained when measurements of
neuronal structure and function are combined. As an example
• A non-invasive imaged-based approach allows quantitative of utility when investigating potential neurotoxic effects,
monitoring of neuronal activity as neurons mature and concentration response curves of paclitaxel were generated
networks develop; the need to select an arbitrary point in time in an established co-culture of primary rat cortical neurons
to perform a single measurement is alleviated. and astrocytes, revealing a substantial reduction in neuronal
activity with only small changes in neurite length after 10 days
• Use of a genetically encoded calcium sensor (IncuCyte of treatment.
NeuroBurst Orange reagent) is non-perturbing and can be
used with a variety of neuronal cell types (such as primary
neurons and iPSC-derived models) to analyze short-term
calcium flux kinetics.

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