Characterization of neurite outgrowth, maturation, and the disruption of neurite networks is key to the study of neuropathological disorders, neuronal injury and regeneration, as well as when screening for neurotoxicity. Additionally, measurements of cell viability are central to the study of drug candidates, culture conditions and the impact of environmental factors on neuronal cell health and function. Neuronal cell health assays are routinely used in drug discovery screens, and may be used to identify treatments that confer neuroprotective effects without causing neurotoxicity.
IncuCyte® Neuronal Cell Health and Morphology Assays reveal both significant and subtle dynamic changes in cells over an extended period of time using fully automated image acquisition and analysis. Our purpose-built software and lab-tested, non-perturbing reagents pair with the IncuCyte® S3 Live-Cell Analysis System for Neuroscience to provide a new, enabling, turnkey solution for analyzing live cells at 96- and 384-well throughput.
IncuCyte® Neuronal Cell Health and Morphology Assays measure cell viability and neurite dynamics, allowing you to:
Generate more data per cell with kinetic assays, and avoid restrictions imposed by end-point analysis.
Reduce artifacts and preserve fragile neurites, with either label-free analysis or use of non-perturbing fluorescent reagents designed for sensitive cells.
Compatible with iPSC-derived or primary cells, in mono- or co-culture.
Easily generate highly reproducible data at microplate throughput suitable for pharmacological analysis.
Examine neurite outgrowth or disruption and cell health with automated, long-term imaging and analysis to capture both significant and subtle dynamic changes in neuronal networks.
Figure 1. Gain valuable biological insight into physiologically relevant in vitro models that more accurately represent human neurodegenerative disease. Quantification of 6-OHDA-mediated toxicity in the Dopa.4U cellular model of Parkinson’s disease (Axol Biosciences). Dopa.4U cells were infected with IncuCyte® NeuroLight Lentiviral Reagent and imaged every 6 hours for 12 days using a 20x objective. 6-OHDA caused a time- and concentration-dependent decrease in neurite length with an IC50 concentration of 84 µM.
Reduce artifacts and preserve fragile neurites. Conduct label-free neurite analysis in mono-culture or use simple, non-perturbing fluorescent labeling in co-culture with IncuCyte NeuroLight Orange Reagent. Detect apoptosis with non-perturbing, mix-and-read IncuCyte® Annexin V Orange Reagent. No fixing, staining, or lifting required.
Figure 2. Perform label-free analysis of neurite length, branch points and cell body clusters automatically and at 96-/384-well throughput. High Definition (HD) phase images of rat cortical neurons are easily segmented using IncuCyte® NeuroTrack Software to identify neurites and cell bodies (A). PlateGraphs show kinetic data in every well, enabling rapid visualization and evaluation of treatment trends in a 384-well screening plate (B).
Figure 3. Analyze neurite dynamics over extended time periods within co-cultures using non-perturbing reagents. Infect neuronal cell types with IncuCyte® NeuroLight Orange Reagent to ensure neuron-specific labeling. Rat cortical neurons cultured in the presence of astrocytes were transduced and imaged over 14 days and treated with glutamate at day 10 (A). Timecourse analysis reveals concentration-dependent treatment effects (B).
Figure 4. Measure time course and concentration-dependent effects on cell viability continuously and noninvasively. IncuCyte® Annexin V Orange Reagent is non-perturbing to cell health and morphology and yields an easily measured fluorescent signal indicative of apoptosis. Observe the kinetics of cell death by analysis of fluorescent signal in images and movies throughout your experiment (A). Analyze the time course of concentration-dependent treatment of neuronal cultures using kinetic readouts (B).
IncuCyte assays and reagents for neurite analysis and cell health are compatible with iPSC-derived or primary cells, in mono- or co-culture.
Figure 5. Examine readouts of neurodegenerative disorders in your preferred primary or iPSC-derived neuronal cell type. IncuCyte® NeuroLight Orange Reagent can be used to selectively label neurites in co-culture for analysis of neurite length. Here, Dopa.4U, hNPC, iCell, and Peri.4U were examined.
Support drug discovery research with data suitable for pharmacological analysis. Evaluate multiple experimental conditions with easy-to-generate, highly reproducible data at microplate throughput. Spend more time investigating, and less-time troubleshooting with our turnkey solution including lab-tested reagents, live-cell protocols, and purpose-built software tools.
Figure 6. Consistent, high volume data sets are rapidly generated through continuous automated image acquisition using minimal cells in 6 x 96- or 6 x 384- well plates in parallel. Time course of iCell Neuron neurite length demonstrating high protocol reproducibility in a 384 well plate over 11 days in culture – CV<15% (A). 384-well plate showing reproducible pharmacology with easily generated EC/IC50 curves (B).
|All Applications for Neuroscience Research|
|IncuCyte® S3 Live-Cell Analysis System for Neuroscience||
Includes image acquisition and analysis system with
|IncuCyte® NeuroTrack Software Module||Enables microplate analysis of neurite dynamics with or without labels||9600-0010|
|IncuCyte® NeuroLight Orange Lentiviral Reagent||Live-cell neuronal labeling reagent for long-term expression of an orange fluorescent protein||4758|
|IncuCyte® NeuroPrime Orange Cell and Reagent Kit||
|IncuCyte® rCortical Neurons||Cryopreserved vial of cortical neurons isolated from embryonic stage (E18) Sprague-Dawley rats (2x106 cells/vial)||4753|
|IncuCyte® rAstrocytes||Cryopreserved vial of astrocytes isolated from embryonic stage (E18) Sprague-Dawley rats (2x106 cells/vial)||4586|
|Neuronal Cell Health|
|IncuCyte® Annexin V Orange Reagent for Apoptosis||One vial of lyophilized reagent sufficient for 100-200 live-cell apoptosis tests in 96-well format||4759|
|IncuCyte® Annexin V NIR Reagent for Apoptosis||One vial of lyophilized reagent sufficient for 100-200 live-cell apoptosis tests in 96-well format||4768|
|IncuCyte® NucLight Orange Lentiviral Reagent||One vial (0.2 mL) for efficient, non-perturbing, nuclear labeling of cells||4771|
Find helpful resources for neuroscience research like peer-reviewed publications, application notes, protocols, scientific posters, videos, and more.
A comprehensive range of reagents and software for the IncuCyte® Live-Cell Analysis System for Neuroscience is available to support your neuroscience research.
Let us know if we can help with your neuroscience research. Request a demo, get a quote, or get general product information from our friendly team.