Neuronal Cell Health & Morphology Overview

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® Live-Cell Analysis System to provide a new, enabling, turnkey solution for analyzing live cells at 96- and 384-well throughput.


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White Paper

Unraveling the Complexities of Neurological Disease and Injury with Real-time Live-cell Analysis

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White Paper

Live-cell Analysis for Neuroscientists: applications for iPSCs and primary cell models

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Key Advantages
  • Gain dynamic insights - Generate more data per cell with kinetic assays, and avoid restrictions imposed by end-point analysis.
  • Protect and preserve your precious cells - Reduce artifacts and preserve fragile neurites, with either label-free analysis or use of non-perturbing fluorescent reagents designed for sensitive cells.  
  • Use your in vitro model of choice - Compatible with iPSC-derived or primary cells, in mono- or co-culture.
  • Get answers faster - Easily generate highly reproducible data at microplate throughput suitable for pharmacological analysis.

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Gain Dynamic Insights

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.

Protect and Preserve your Precious Cells

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).

Use your in vitro Model of Choice

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.

Get Answers Faster

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).

Assays

Learn More about Assays for Neuronal Cell Health and Morphology

Incucyte Neurite Outgrowth Assays

Key Advantages:

  • Generate kinetic, image-based and automated measurements using Incucyte Neurotrack Software Module
  • Capture subtle, transient events often missed with end-point assays – Incucyte enables non-invasive, repeated measurements of the same neuronal network
  • Generate more information-rich data using simple, cell sparing protocols - no cell lifting or fixing/staining


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Incucyte® Annexin V Orange Dye for Apoptosis

Key Advantages:

  • Real-time, automated analysis of the time course of apoptosis suitable for pharmacology studies
  • Visualize morphological changes and validate treatment effects with images and movies
  • Maintain cell health and morphology while reducing loss of precious cells with a non-perturbing, mix-and-read reagent


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Ordering Information

Application

Description

Cat. No.

Instrument

Incucyte® SX5 Live-Cell Analysis System

Our newest and most advanced offering, supporting more applications and up to 5 fluorescence channels

4816

Neurite Dynamics

Incucyte® NeuroTrack Software Module

Enables microplate analysis of neurite dynamics with or without labels

9600-0010

Incucyte® NeuroLight Orange Lentivirus

Live-cell neuronal labeling reagent for long-term expression of an orange fluorescent protein

4808

Incucyte® NeuroLight Red Lentivirus

Live-cell neuronal labeling reagent for long-term expression of a red fluorescent protein

4807

Incucyte® NeuroPrime Orange Kit

Kit includes:

  • Incucyte® NeuroLight Orange Lentivirus Reagent
  • Incucyte® rAstrocytes
  • Incucyte® rCortical Neurons

4760

Incucyte® NeuroPrime Red Kit

Kit includes:

  • Incucyte® NeuroLight Red Lentivirus Reagent
  • Incucyte® rAstrocytes
  • Incucyte® rCortical Neurons

4585

Neuronal Cell Health

Incucyte® Annexin V Green Reagent

One vial of lyophilized reagent sufficient for 100-200 live-cell apoptosis tests in 96-well format

4642

Incucyte® Annexin V Orange Reagent

One vial of lyophilized reagent sufficient for 100-200 live-cell apoptosis tests in 96-well format

4759

Incucyte® Annexin V Red Reagent

One vial of lyophilized reagent sufficient for 100-200 live-cell apoptosis tests in 96-well format

4641

Incucyte® Annexin V NIR Reagent

One vial of lyophilized reagent sufficient for 100-200 live-cell apoptosis tests in 96-well format

4768

Cell Lines

Incucyte® rCortical Neurons

One cryopreserved vial of rat cortical neurons at a density of 2x106 cells/vial

4753

Incucyte® rAstrocytes

One cryopreserved vial of rat astrocytes at a density of 2x106 cells/vial    

4586


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Resources and Applications

Resources

Protocols

Incucyte® Annexin V NIR and Incucyte® NeuroLight Orange Multiplex

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Neuroscience Literature & Resources

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Neuroscience Reagents & Software

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