Neuronal Networks

 

Primary Cell Cultures on MaxOne MEA

Dissociated cell cultures can be plated, grown, and imaged on MaxOne MEAs. The figure shows an overlay of multiple confocal stack images: cell bodies and neurites are visible on the left side and the electrode array is focused on the right side.

Protocol

Sample Cell Culture Plating Procedure

  1. Pre-coat the electrode array surface with a thin layer of poly(ethyleneimine) (Sigma, Missouri, USA), 0.05% by weight in borate buffer (Chemie Brunschwig, Basel, Switzerland) at 8.5 pH.
  2. Apply a 10 μl drop of 0.02 mg/ml laminin (Sigma) in Neurobasal medium (Invitrogen, California, USA) for cell adhesion.
  3. Seed cell suspension in a 6-μl drop over the array.
  4. Add 1 ml of plating medium after 20-30 minutes.
  5. After 24 hours, the plating medium was changed to growth medium.
  6. Maintain the cultures inside an incubator to control environmental conditions (37 °C, 65% humidity, 5% CO2)
  7. Use 1-2 ml of growth medium. Replace 50% of the medium twice per week.

Sample Cell Culture Medium

  • Plating medium consists of 850 ml of Neurobasal, supplemented with 10% horse serum (HyClone, Utah, USA), 0.5 mM GlutaMAX (Invitrogen, California, USA) and 2% B27 (Invitrogen, California, USA).
  • Growth medium consists of 850 ml of DMEM (Invitrogen, California, USA), supplemented with 10% horse serum, 0.5 mM GlutaMAX and 1 mM sodium pyruvate (Invitrogen, California, USA).

Single Cell Level Analysis

Isolate extracellular action potentials from individual cells easily.

Select electrodes with the best signals from each cell and improve your spike sorting.

Analyze subcellular features, such as full axonal arbors in single neurons.

Electrical signals tracked along neurites enable to investigate novel parameters, difficult to obtain with previous technologies.

Delay map

Amplitude map

High-resolution axonal action potential propagation tracking


 

 

The video shows the propagation of an AP through the axonal arbor of a single neuron. Electrical signals tracked along neurites enable to investigate novel parameters, difficult to obtain with previous technologies.

 

Tracking axonal branch signals


 

 

The video shows the propagation of a stimulation-induced action potential through a branching segment of an axon. To reduce noise and better reveal the action potential, 300 trials have been averaged.

 

Network Level Analysis

Record and/or stimulate every cell on the MEA.

MaxOne allows simultaneous recording from thousands of cells yields rich datasets for studying neuronal networks. Stimulation experiments can be designed with high flexibility to manipulate network dynamics.

Electrically image active cells on the MEA.

Localize active cells on the MEA during experiments. The optical image from fluorescence microscopy matches the electrical image obtained using MaxOne.


Electrical image

 

Study cells and networks across different spatial and temporal scales.

Action potentials and local field potentials can be simultaneously recorded by MaxOne over time scales of microseconds to months. The raster plot shows the dynamics of network activity using 1,024 active electrode sites.

Raster plot

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