Neurite Outgrowth — Quantifying Neurite Dynamics in Real Time

During development, neurons extend numerous processes that differentiate into dendrites and axons. These processes (termed neurites) are critical for communication between neurons. Characterization of neurite outgrowth, maturation, and the disruption of neurite networks is key to the study of neuropathological disorders (e.g. neurodegeneration), neuronal injury/regeneration and screening for neurotoxicity. However, current techniques used to study those dynamic biological changes are challenging.

With the IncuCyte® Live-Cell Analysis System and IncuCyte® NeuroTrack Software Module (Cat. No. 9600-0010) you can now achieve automated, continuous, non-invasive imaging and measurement of neurite outgrowth with 96-/384-well assays that generate meaningful, kinetic data with visual confirmation of biological changes.


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Key Features and Benefits

Feature

Benefit

  • Unique, mobile optical train keeps cells stationary while optics move
  • Incubation is provided by your tissue culture incubator
Conduct long-term study (weeks or months) on sensitive cells that are physically undisturbed and in a consistent, physiologically-relevant environment. Avoid damage to delicate neurite structures.
Automated image acquisition and analysis enables kinetic measurements in 96-well formats, up to six microplates in parallelConduct robust pharmacological analysis or study more variables in less time
Purpose-built, guided image analysis workflow measures neurite structures and branch points in high-contrast phase imagesEasily quantify neurite outgrowth, label-free 
Proprietary reagents fluorescently-label neurites without perturbing cell health or morphologyPerform kinetic neurite outgrowth analysis in co-cultures

Simple, cell sparing protocols without fixing, staining, or antibody-labeling

Generate more information-rich data using less of your precious samples
Complete end-to-end solution including instrument, software and reagentsReduce time spent troubleshooting and spend more time investigating
Neurite Analysis Assays

Real-Time Neurite Analysis Inside Your Incubator

IncuCyte® Neurite Analysis Assays enable automated, continuous analysis of neurite outgrowth using your choice of neurons (e.g. primary, iPSC-derived, immortalized neuronal cell lines) in monoculture or in co-culture with astrocytes. Quantify neurite length and branch points in order to evaluate neural network stability over time – all inside your cell culture incubator.

  • Screen for neurotoxicity
  • Investigate neuroprotective treatments
  • Study pathways involved in neurodegeneration
  • Measure neuronal differentiation in real time

1. Label-Free Analysis of Neurons Inmonoculture

Monitor neurite outgrowth in live neuronal monocultures using IncuCyte NeuroTrack analysis software. Fully automated, long-term label-free analysis. Human iPSC-derived neurons (iCell Neurons, CDI) in monoculture.

2. Fluorescence Analysis of Neurons in Co-Culture With Astrocytes

Measure changes in neurite length and branch points in mixed cultures of neurons and astrocytes using fluorescent neuronal labelling reagents and IncuCyte analysis software.  Primary rat forebrain neurons labeled with the IncuCyte® NeuroLight Orange reagent in co-culture with rat astrocytes.

Key Advantages

Gain Dynamic, Relevant Insight

Visualize and quantify long-term changes in neurite outgrowth and disruption in real time in label-free monocultures or use IncuCyte® NeuroLight reagent or NeuroPrime Cell Kits for selective analysis of neurite dynamics in co-cultures.


Figure 1. Continuous measures of neurite dynamics.. IncuCyte NeuroTrack analysis software automatically analyzes images to provide measures of neurite length, branch points and neuronal cell body clusters.


Maximize Your Neuroscience Research

Our turnkey solution enables automatic generation of time courses to reveal concentration-dependent responses with IncuCyte NeuroTrack’s unbiased, automated analysis in 96- or 384-well plates.


Figure 2. Measure treatment effects automatically and non-invasively. IncuCyte neurite analysis assays allow every well of a 96- or 384-well plate to be imaged and analyzed automatically to provide a microplate readout of neurite dynamics over time (A). Time-courses reveal concentration-dependent treatment effects (B). Transform data into concentration-response curves to compare pharmacology (C).

Use Your in Vitro Model of Choice Neurodegeneration and Neurotoxicity

Make continuous measurements in relevant neuronal models in highly flexible, robust and reproducible assay formats suitable for screening and profiling. IncuCyte assays and reagents are compatible with iPSC-derived or primary cells, in mono- or co-culture.


Figure 3. Create relevant in vitro models of neurodegeneration and neurotoxicity — screen for neuroprotective treatments. Concentration dependent neurotoxic effects of 6-hydroxydopamine (6-OHDA) measured in human iPSC-derived dopaminergic neurons (Dopa.4U, Axiogenesis) in a functional model of Parkinson’s disease. Treatment with bone derived neural factor (BDNF) promotes neurite formation but provided little or no protection against the effects of 6-OHDA.

Protect Your Cells

Real- time live cell protocols for continuous measurements of neuronal networks in monocultures or co-cultures using IncuCyte NeuroLight Lentiviral labeling reagents – no fixing, washing or labeling steps!

Neuronal monocultures—label-free protocol overview

Neuronal co-cultures—fluorescence protocol overview

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Learn how IncuCyte live-cell analysis has been used by researchers around the world to drive neuroscience discovery.

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Ordering Information

Ordering Information

Product

Qty.

Cat. No.

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

IncuCyte® rCortical Neurons2 x 106 cells/vial

4753

IncuCyte® rAstrocytes2 x 106 cells/vial

4586

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


Resources

Literature and Documentation

Product Data Sheet

IncuCyte® NeuroLight Orange Lentivirus—Synapsin Promoter

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Protocol: Label-free Neurite Analysis

View the Protocol

Protocol: Fluorescence Neurite Analysis

View the Protocol

Product Guide: IncuCyte® NeuroPrime Kit

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SFN 2014 Poster

Kinetic, label-free assays for neurite outgrowth in primary, iPSC-derived and immortalized neurons

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