Functional imaging of neuron–astrocyte interactions in a compartmentalized microfluidic device

Traditional approaches in cultivating neural cells in a dish without orienting their interactions have had only limited success in revealing neural network properties. To enhance the experimental capabilities of studying neural circuitry in vitro, we designed an experimental system combining concepts of micropatterned surfaces, microfluidic devices and genetically encoded biosensors. Micropatterning was used to position neurons and astrocytes in defined locations and guide interactions between the two cell types. Microfluidic chambers were placed atop micropatterned surfaces to allow delivery of different pharmacological agents or viral vectors to the desired cell types. In this device, astrocytes and neurons communicated through grooves molded into the floor of the microfluidic device. By combining microfluidics with genetically encoded calcium indicators as functional readouts, we further demonstrated the utility of this device for analyzing neuron–neuron and neuron–astrocyte interactions in vitro under both healthy and pathophysiological conditions. We found that both spontaneous and evoked calcium dynamics in astrocytes can be modulated by interactions with neurons. In the future, we foresee employing the microdevices described here for studying mechanisms of neurological disorders.