AxonTracking Assay

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AxonTracking Assay

A Novel Approach

Neurons communicate within a network via action potentials (APs) propagating along axons. The capability to  access axonal physiology is crucial for studying information processing among neurons in healthy and diseased states. However, axonal signals are difficult to measure at a large-scale. Therefore, the combination of reliability, ease of use, throughput, long-term and non-invasive measurement are necessary to monitor and understand neuronal function at a scale that was previously not possible.

High-density microelectrode array (HD-MEA) measurements at unprecedented resolution and high signal quality, using MaxOne and MaxTwo systems, allow to detect the AP propagation from the initiation site down to distal axonal branches. With the AxonTracking Assay, the identification of the axonal paths is fully automated at the micrometer scale. This live-cell recording and analysis provides novel functional and structural readouts applicable for phenotypic characterization, disease modeling, and drug screening studies.

 

Automated

Long-Term

Label-Free

The fully automated platform is easy to use and
allows for simultaneous recordings of multiple
neurons and axonal branches in multiple wells.
Characterize neuronal maturation, development
or treatment effects by recording from your
culture over multiple days and weeks.
The electrical recordings are noninvasive and
label-free, which avoids introducing side effects
from dyes etc.

 

HD-MEA Technology for Recording Axonal Signals

Powered by MaxOne and MaxTwo

Key advantages:

High Spatio-Temporal Resolution
Reconstruct axonal paths by tracking Action Potential propagation at
thousands of sites, thanks to the densely packed microelectrode array.
Large Sensor Area
Detect long axonal branches of multiple neurons at the same time with
a large sensor area, applicable for 2D and 3D samples.
High Signal Quality
Catch the smallest signals propagating along axons, down to single
micro-volts-range, with low-noise recording channels

Experimental Workflow

 

 

 

 

 

 

 

Assay Workflow





Record


Record the active neurons
identified with the ActivityScan
Assay.




Process


Reveal the axonal morphologies
through a series of processing steps:
◉ Spike sorting
◉ Spike-triggered averaging
◉ Footprint extraction




Analyze


Identify individual axonal branches
and reconstruct the morphology of the
neurite outgrowth using an unsuper- vised object-tracking algorithm.

Metrics

Legend

1 – Neuron Conduction Velocity
2 – Total Detected Axon Length
3 – Longest Branch Length
4 – Longest Latency
5 – Longest Distance from Initiation Site
6 – Amplitude at Initiation Site

Results

AxonTracking Assay in Human Neurons

Propagating APs along axonal processes recorded from different human induced pluripotent stem cells-derived (iPSC-derived) neuronal cell lines.

Legend

1 – Human Glutamatergic Neurons
2 – Human Dopaminergic Neurons
3 – Human Motor Neurons

AxonTracking Assay in Long-Term Neuronal Cultures

Neurons and propagating APs can be resolved even in cultures with long and dense axonal processes (iPSC derived glutamatergic neurons, DIV 63)

Legend

1 – Activity map
2 – AxonTracking Assay
3 – Propagating AP of individual neurons resolved

 

AP Propagation Along Axons

Propagating APs along the axonal processes of a rat primary cortical neuron shown in a time-series:

References

Bakkum, D. J., Frey, U., Radivojevic, M., Russell, T. L., Müller, J., Fiscella, M., Takahashi, H., & Hierlemann, A. “Tracking axonal action potential propagation on a high-density microelectrode array across hundreds of sites.” Nat Commun. 4, 2181 (2013).

Bullmann, T., Radivojevic, M., Huber, S. T., Deligkaris, K., Hierlemann, A., & Frey, U. “Large Scale Mapping of Axonal Arbors Using High-Density Microelectrode Arrays.” Front. Cellular Neurosci. 13, 404 (2019).

 

AxonTracking Assay Brochure

AxonTracking Assay Brochure | Download Here 

 

Want to try this with your cells? Contact us today

We would love to answer any of your questions! Book a one-to-one call with one of our scientist to discuss how MaxWell Biosystems’ high-content electrophysiology solutions can bring new key insights to your project or request a quote.

Track your Axon to WIN!

Do you have a cool image of a tracked neuron using our AxonTracking Assay? Share your image and you could win a $2000 Travel Grant to SfN 2022 in San Diego!

Find out how to enter the competition.

 

 

Page updated 03/08/2022

 

 

 

Contacts






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8047 Zurich, Switzerland
+41 44 244 24 24
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info@mxwbio.com
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