Neuronal Cell Health & Morphology Overview

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 Lentivirus 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 Lentivirus. Detect apoptosis with non-perturbing, mix-and-read Incucyte^® Annexin V Orange Dye. 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 Module  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 Lentivirus 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 Dye 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 Lentivirus*** 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).