Reconstruction of directed neuronal networks in a microfluidic device with asymmetric microchannels.

Microfluidic devices for controlling neuronal connectivity in vitro are extremely useful tools for deciphering pathological and physiological processes occurring in neuronal networks. These devices allow the connection between different neuronal populations located into separate culture chambers through axon-selective microchannels. In order to implement specific features of brain connectivity such as directionality, it is necessary to control axonal growth orientation in these devices. Among the various strategies proposed to achieve this goal, one of the most promising and easily reproducible is the use of asymmetric microchannels. We present here a general protocol and several guidelines for the design, production and testing of a new paradigm of asymmetric microchannels geometries based on a "return to sender" strategy. In this method, axons are either allowed to travel between the emitting and receiving chambers within straight microchannels (forward direction), or are rerouted toward their initial location through curved microchannels (reverse direction). We introduce variations of these "arches" microchannels and evaluate their respective axonal filtering capacities. Importantly, one of these variants presents an almost complete filtration of axonal growth in the non-permissive direction while allowing robust axonal invasion in the other one, with a selectivity ratio as high as 99.7%.

Transient microfluidic compartmentalization using actionable microfilaments for biochemical assays, cell culture and organs-on-chip.

We report here a simple yet robust transient compartmentalization system for microfluidic platforms. Cylindrical microfilaments made of commercially available fishing lines are embedded in a microfluidic chamber and employed as removable walls, dividing the chamber into several compartments. These partitions allow tight sealing for hours, and can be removed at any time by longitudinal sliding with minimal hydrodynamic perturbation. This allows the easy implementation of various functions, previously impossible or requiring more complex instrumentation. In this study, we demonstrate the applications of our strategy, firstly to trigger chemical diffusion, then to make surface co-coating or cell co-culture on a two-dimensional substrate, and finally to form multiple cell-laden hydrogel compartments for three-dimensional cell co-culture in a microfluidic device. This technology provides easy and low-cost solutions, without the use of pneumatic valves or external equipment, for constructing well-controlled microenvironments for biochemical and cellular assays.

Effect of threshold disorder on the quorum percolation model.

We study the modifications induced in the behavior of the quorum percolation model on neural networks with Gaussian in-degree by taking into account an uncorrelated Gaussian thresholds variability. We derive a mean-field approach and show its relevance by carrying out explicit Monte Carlo simulations. It turns out that such a disorder shifts the position of the percolation transition, impacts the size of the giant cluster, and can even destroy the transition. Moreover, we highlight the occurrence of disorder independent fixed points above the quorum critical value. The mean-field approach enables us to interpret these effects in terms of activation probability. A finite-size analysis enables us to show that the order parameter is weakly self-averaging with an exponent independent on the thresholds disorder. Last, we show that the effects of the thresholds and connectivity disorders cannot be easily discriminated from the measured averaged physical quantities.

Asymmetric axonal edge guidance: a new paradigm for building oriented neuronal networks.

We present a novel kind of directional axon guides for brain-on-a-chip applications. Contrarily to previous works, the directionality in our design is created by rerouting axons growing in the unwanted direction back to their original compartment while leaving the other growth direction unaffected. This design yields state-of-the-art levels of directionality without the disadvantages of previously reported technologies.

Combining microfluidics, optogenetics and calcium imaging to study neuronal communication in vitro.

In this paper we report the combination of microfluidics, optogenetics and calcium imaging as a cheap and convenient platform to study synaptic communication between neuronal populations in vitro. We first show that Calcium Orange indicator is compatible in vitro with a commonly used Channelrhodopsine-2 (ChR2) variant, as standard calcium imaging conditions did not alter significantly the activity of transduced cultures of rodent primary neurons. A fast, robust and scalable process for micro-chip fabrication was developed in parallel to build micro-compartmented cultures. Coupling optical fibers to each micro-compartment allowed for the independent control of ChR2 activation in the different populations without crosstalk. By analyzing the post-stimuli activity across the different populations, we finally show how this platform can be used to evaluate quantitatively the effective connectivity between connected neuronal populations.

Memory decay and loss of criticality in quorum percolation.

In this paper, we present the effects of memory decay on a bootstrap percolation model applied to random directed graphs (quorum percolation). The addition of decay was motivated by its natural occurrence in physical systems previously described by percolation theory, such as cultured neuronal networks, where decay originates from ionic leakage through the membrane of neurons and/or synaptic depression. Surprisingly, this feature alone appears to change the critical behavior of the percolation transition, where discontinuities are replaced by steep but finite slopes. Using different numerical approaches, we show evidence for this qualitative change even for very small decay values. In experiments where the steepest slopes can not be resolved and still appear as discontinuities, decay produces nonetheless a quantitative difference on the location of the apparent critical point. We discuss how this shift impacts network connectivity previously estimated without considering decay. In addition to this particular example, we believe that other percolation models are worth reinvestigating, taking into account similar sorts of memory decay.