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Functional phenotyping is the quantitative assessment of cellular activity, particularly electrophysiological function, used to characterize cell behavior beyond morphology or molecular and genetic identity. In neuroscience and stem cell biology, it involves capturing the dynamics of neuronal function, including how neurons generate and propagate action potentials, form synaptic connections, and engage in coordinated network activity over time. By measuring both spontaneous and evoked activity at single-cell and network levels, functional phenotyping provides critical insights into neuronal maturation, connectivity, and physiological relevance. These dynamic functional readouts complement imaging and transcriptomic profiling, offering a more comprehensive understanding of neuronal identity and behavior.
MaxWell Biosystems’ HD-MEA platforms offer a powerful approach to functional phenotyping, capturing rich, multidimensional data beyond basic firing rates. With ultra high-resolution, label-free recordings of neuronal activity from subcellular events to large-scale networks, our HD-MEAs allow scientists to reveal the full story encoded in their biological models.
Network bursting activity provides insights into neuronal maturation and connectivity. Reproducibility across wells, batches, and timepoints gives researchers confidence that their observations reflect true biology, not technical noise or variability caused by limited coverage in each well.
Different cell types and maturation stages show distinct axonal morphologies and conduction velocities. Detailed axon tracking enriches cell classification and enables researchers to assess how efficiently and precisely neurons communicate.
Early-stage neuronal cultures and 3D models like organoids often exhibit weak, sparse signals. Detecting these subtle events is key to tracking the onset of excitability, connectivity, and functional maturation. Reliable detection of low-amplitude activity also supports comprehensive, longitudinal tracking of functional changes.
For large-scale studies with diverse cell lines and models, automation ensures consistent data acquisition, processing, and analysis across wells and timepoints. Our automation-ready HD-MEAs streamline workflows, accelerates data turnaround, and enables scientists to quickly evaluate and interpret phenotypes.
In this study, HD-MEA technology was used to characterize and compare the functional development of human iPSC-derived motor and dopaminergic neurons (iCell® DopaNeurons, iCell® Motor Neurons, iCell® Astrocytes, FUJIFILM Cellular Dynamics International, Madison, USA). Cultures were recorded at multiple time points (DIV 7, 14, and 21), capturing the maturation of spontaneous firing and network activity over time. Using Activity Scan and Network Metrics, researchers were able to distinguish between cell lines based on their electrophysiological phenotypes. The study also included comparisons of isogenic lines modeling Parkinson’s disease and ALS, with additional insights from Axon Tracking analysis. Read more in Ronchi et al., 2021.
Electrophysiological phenotype characterization of human iPSC-derived neurons across development
Top: 2D spatial distribution maps of firing rates for motor neurons (left) and dopaminergic neurons (right) at DIVs 7, 14, 21, respectively.
Bottom: Network activity, depicted as population spike time histograms, of motor neurons (left) and dopaminergic neurons (right) at DIVs 7, 14, 21, respectively.
Data adapted from Ronchi et al., Advanced Biology, 2021
This case study demonstrates axonal development and maturation over time in SynFire® iN co-cultures with astrocytes, utilizing MaxWell Biosystems’ Axon Tracking Assay. The data reveal dynamic structural and functional changes, underscoring the assay’s sensitivity to long-term neuronal growth.
The developmental process of axonal formation and maturation captured over time using the MaxTwo HD-MEA system.
First row: Tracked axonal trajectories (red contours) overlaid on electrical activity footprints obtained from multiple neurons within the network. A notable increase in both structural and functional complexity of the axons is observed when comparing data at DIV 21 (Left) and DIV 42 (Right).
Second, Third, and Fourth rows: Electrical footprints of selected neurons from the same well recorded at DIV 21 (Left) and DIV 42 (Right). Quantitative, single-cell metrics, such as conduction velocity, were automatically extracted using MaxLab Live, uncovering clear, time-dependent development of subcellular features that reflect the functional maturation of the neuronal network.
This case study presents a comprehensive and multifaceted functional profile of neuronal activity across diverse cell lines, examining subcellular, cellular, and network-level dynamics. The full spectrum of functional profiles reveals significant differences among various neuronal cultures at multiple levels of detail.
From left to right: Heatmaps display the spatial distribution of spiking activity across the culture; raster plots illustrate the timing and coordination of population bursts; and axonal footprint reconstructions uncover the subcellular paths of signal propagation for individual neurons. With MaxLab Live, you can quantify these features across multiple scales (subcellular, single-cell, and network), revealing functional phenotypes that would otherwise remain hidden.
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