Neurite Outgrowth

Gain valuable insights for interpreting neurobiology, from evaluation of iPSC-derived neurons to analysis of 3D neuron organoids.

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Simplify characterization of neurons using neurite outgrowth assay to study neuronal development and degeneration 𝘪𝘯 𝘷𝘪𝘵𝘳𝘰

Neurons create connections via extensions of their cellular body called axons and dendrites, which are commonly referred to as “neurites” or “processes”. This biological phenomenon is referred to as neurite outgrowth and is regulated by complex intracellular signaling events.

Neurite outgrowth is a commonly used assay to study neuronal development and neuronal degeneration in vitro. Development of neurites requires a complex interplay of both extracellular and intracellular signals. The growth of neurites can be stimulated or inhibited by neurotrophic factors. Importantly, the development of neurons can be affected by neurotoxic chemicals.

Understanding the signaling mechanisms driving neurite outgrowth provides valuable insight for interpreting neurotoxic responses and compound screening data and for interpreting factors influencing neural development and regeneration. Inhibition or stimulation of neurite outgrowth is implicated in a broad range of CNS disorders or injuries including stroke, Parkinson’s disease, Alzheimer’s disease, and spinal cord injuries.

Workflow solution for analyzing neurite outgrowth

Neurite outgrowth is assessed by the segmentation and quantification of neuronal processes. These neuronal processes can be imaged using a fluorescence microscope and quantified with manual tracing and counting when throughput is low. However, for samples in a higher-throughput microplate format, an automated imaging system paired with analysis software is a more efficient solution.

Neurite Workflow

The workflow illustrates a simplified process for analyzing neurite outgrowth and highlights systems to help you streamline your research and increase your throughput.

  1. Culture neuronal cells – cells were grown and allowed to form neurite networks in 96- or 384-well microplates.
  2. Treat with compounds – the cells were then exposed to toxic compounds for 48 hours.
  3. Stain for markers – After compound treatment is complete, live cell stains can be added directly to the media. Immunostaining protocols with fluorescently-conjugated antibodies can also be performed post-cell fixation.
  4. Acquire neuronal images – High-content imaging of neurons allows scientists to both characterize and measure changes in neuronal networks such as neurite number, length, and branching, as well as to determine gross or specific toxicity reactions. Acquire images with large field-of-view optics so more cells can be sampled with fewer sites per well, leading to dramatically faster plate acquisition times.
  5. Analyze neuronal network – High content analysis provides a quantitative method to determine effects of positive and negative factors on neurite outgrowth. Use cellular imaging analysis software to run quantitative analysis of the neuronal cell images to characterize several parameters including number of processes per cell, length of neurite outgrowth, branching, and number of cells.

Neurite outgrowth applications and assays

High-content imaging of neurons allows scientists to both characterize and measure changes in neuronal networks such as neurite number, length, and branching, as well as to determine gross or specific toxicity reactions.

Learn how to capture and quantify neuronal activities quickly and accurately using automated microscopy and high-content analysis software:

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