IncuCyte Research Areas
Neurodegenerative diseases are characterized by the progressive loss of structure and function of cells comprising the nervous system. Because subtle, progressive changes contributing to disease etiology are difficult to capture, characterizing these dynamic processes requires techniques that consistently monitor cell biology in vitro over time.
Unlike other technologies, such as High Content Imaging and time-lapse microscopy, the IncuCyte® S3 Live-Cell Analysis System allows you to capture small, dynamic changes in cells over an extended period of time using fully automated image acquisition and analysis. Collect kinetic measurements of cell health, structure, function, and neurite disruption in vitro with uninterrupted incubation provided by a standard cell culture incubator. Non-perturbing reagents preserve cell morphology without the disruption and restrictions imposed by end-point analysis, thus allowing more data collection from every cell.
The IncuCyte S3 Live-Cell Analysis System, Neurite Analysis software module, and IncuCyte reagents and cell kits provide a new, enabling, and end-to-end solution for analyzing live cells with 96- and 384-well throughput.
Gain dynamic insight into mechanisms of neurodegeneration and neuroprotection. See what you can do with IncuCyte S3 Live-Cell Analysis System:
Explore disease models with long term, kinetic, image-based measurements.
Quantify dynamic changes in neurite outgrowth and cell health
Preserve precious samples with multiplexed, long-term monitoring
|・ 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 parallel||Conduct 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 images||Easily quantify neurite outgrowth, label-free|
|Proprietary reagents fluorescently label neurites or detect cell health without perturbing cell health or morphology||Perform kinetic neurite outgrowth analysis in co-cultures, and multiplex with cell health readouts for additional insight|
|Simple, cell sparing protocols without fixing, staining, or antibody-labelling||Generate more information-rich data using less of your precious samples|
|Complete end-to-end solution including instrument, software, reagents, and cell kits||Reduce time spent troubleshooting and spend more time investigating|
Figure 1. Gain valuable biological insight into physiologically relevant in vitro models that more accurately represent human neurodegenerative disease. Fluorescent NeuroTrack images and quantification of 6-OHDA-mediated toxicity in the Dopa.4U cellular model of Parkinson’s disease (Ncardia). Dopa.4U cells were infected with IncuCyte NeuroLight Red 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.
Figure 2. Discover how long-term, kinetic, label-free measurements of neurite length using IncuCyte Neurotrack Software can reveal differences in drug-induced structural plasticity during neurodegeneration and neuroprotection. Kinetic time course of alterations in neurite length of iCell Neurons (CDI) with kainate (KA) alone or in combination with the AMPA receptor blocker CNQX or the growth factor BDNF (left panel). CNQX completely blocks kainate-induced neurotoxicity, while BDNF treatment results in neurite recovery at later time points. In contrast to expectation, when challenged with KA, we observed no neuro-protective effect with H1152, where a greater loss of neurites was seen at late time-points. Comparison of % neuroprotection of modulators of neurite outgrowth between late and early phase time points (right panel).
Figure 3. Multiplex neurite outgrowth analysis with cell health measurements in physiologically relevant human iPSC-derived neuronal models.
(Top figure) Representative time-lapse movies of rat forebrain neurons in coculture with rat astrocytes. Cells were infected with IncuCyte NeuroLight Red Reagent and incubated with IncuCyte Annexin V Green reagent to monitor neurite length and apoptosis, respectively. Movies were generated over 72 h following treatment with vehicle (left) or 1mM kainate (right).
(Lower figures) Time course of the effects of kainate addition on neurite length (A) and cell viability (B) of iCell GlutaNeurons in co-culture with rat astrocytes. Kainate caused a concentration-dependent decrease in neurite length with concomitant decrease in cell viability (increased IncuCyte Annexin V Green fluorescence). Addition of AMPA receptor antagonist NBQX protects iCell GlutaNeurons from excitotoxicity.
Figure 4. Multiplex neurite outgrowth and cell health measurements in primary neuronal cultures to monitor well-established readouts for studying neurodegeneration. Time course of the effects of glutamate addition at day 8 of culture on neurite length (left panel) and cell viability (middle panel) of primary rat forebrain neurons in co-culture with rat astrocytes. Glutamate treatment caused a concentration-dependent decrease in neurite length with concomitant decrease in cell viability (increased IncuCyte Annexin V Green fluorescence). Glutamate concentration legend is the same for both readouts. Single time-point analysis done at peak response to glutamate (right panel) – Neurite Length (24 hrs post-treatment) - IC50 = 16 µM; Apoptotic cell death (8 hrs post-treatment) EC50 = 21 µM (Annexin V Green).
Figure 5. Examine readouts of neurodegenerative disorders in your preferred primary or iPSC-derived neuronal cell type. 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. IncuCyte NeuroLight Red reagent can be used to monitor neurite length in a variety of experimental cell models (top figures). Time course of iCell Neuron neurite length demonstrating high protocol reproducibility in a 384 well plate over 11 days in culture – CV<15% (bottom left figure and insert). 384 well plate showing reproducible pharmacology with easily generated EC/IC50 curves (bottom right figure and insert).
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 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.