Comparison of fractal and grid electrodes for studying the effects of spatial confinement on dissociated retinal neuronal and glial behavior.

Understanding the impact of the geometry and material composition of electrodes on the survival and behavior of retinal cells is of importance for both fundamental cell studies and neuromodulation applications. We investigate how dissociated retinal cells from C57BL/6J mice interact with electrodes made of vertically-aligned carbon nanotubes grown on silicon dioxide substrates. We compare electrodes with different degrees of spatial confinement, specifically fractal and grid electrodes featuring connected and disconnected gaps between the electrodes, respectively. For both electrodes, we find that neuron processes predominantly accumulate on the electrode rather than the gap surfaces and that this behavior is strongest for the grid electrodes. However, the 'closed' character of the grid electrode gaps inhibits glia from covering the gap surfaces. This lack of glial coverage for the grids is expected to have long-term detrimental effects on neuronal survival and electrical activity. In contrast, the interconnected gaps within the fractal electrodes promote glial coverage. We describe the differing cell responses to the two electrodes and hypothesize that there is an optimal geometry that maximizes the positive response of both neurons and glia when interacting with electrodes.

Controlled assembly of retinal cells on fractal and Euclidean electrodes.

Controlled assembly of retinal cells on artificial surfaces is important for fundamental cell research and medical applications. We investigate fractal electrodes with branches of vertically-aligned carbon nanotubes and silicon dioxide gaps between the branches that form repeating patterns spanning from micro- to milli-meters, along with single-scaled Euclidean electrodes. Fluorescence and electron microscopy show neurons adhere in large numbers to branches while glial cells cover the gaps. This ensures neurons will be close to the electrodes' stimulating electric fields in applications. Furthermore, glia won't hinder neuron-branch interactions but will be sufficiently close for neurons to benefit from the glia's life-supporting functions. This cell 'herding' is adjusted using the fractal electrode's dimension and number of repeating levels. We explain how this tuning facilitates substantial glial coverage in the gaps which fuels neural networks with small-world structural characteristics. The large branch-gap interface then allows these networks to connect to the neuron-rich branches.

Investigation of Fractal Carbon Nanotube Networks for Biophilic Neural Sensing Applications.

We propose a carbon-nanotube-based neural sensor designed to exploit the electrical sensitivity of an inhomogeneous fractal network of conducting channels. This network forms the active layer of a multi-electrode field effect transistor that in future applications will be gated by the electrical potential associated with neuronal signals. Using a combination of simulated and fabricated networks, we show that thin films of randomly-arranged carbon nanotubes (CNTs) self-assemble into a network featuring statistical fractal characteristics. The extent to which the network's non-linear responses will generate a superior detection of the neuron's signal is expected to depend on both the CNT electrical properties and the geometric properties of the assembled network. We therefore perform exploratory experiments that use metallic gates to mimic the potentials generated by neurons. We demonstrate that the fractal scaling properties of the network, along with their intrinsic asymmetry, generate electrical signatures that depend on the potential's location. We discuss how these properties can be exploited for future neural sensors.

Droplet-based high-throughput cultivation for accurate screening of antibiotic resistant gut microbes.

Traditional cultivation approaches in microbiology are labor-intensive, low-throughput, and yield biased sampling of environmental microbes due to ecological and evolutionary factors. New strategies are needed for ample representation of rare taxa and slow-growers that are often outcompeted by fast-growers in cultivation experiments. Here we describe a microfluidic platform that anaerobically isolates and cultivates microbial cells in millions of picoliter droplets and automatically sorts them based on colony density to enhance slow-growing organisms. We applied our strategy to a fecal microbiota transplant (FMT) donor stool using multiple growth media, and found significant increase in taxonomic richness and larger representation of rare and clinically relevant taxa among droplet-grown cells compared to conventional plates. Furthermore, screening the FMT donor stool for antibiotic resistance revealed 21 populations that evaded detection in plate-based assessment of antibiotic resistance. Our method improves cultivation-based surveys of diverse microbiomes to gain deeper insights into microbial functioning and lifestyles.

The human gut is inhabited with hundreds of billions of bacterial cells from a wide range of families. This complex mixture of bacteria is part of the gut microbiome, along with other lifeforms such as viruses, archaea and fungi. As well as interacting with each other, the bacteria in the microbiome interact with our cells and available nutrients. Studying these interactions can help us understand how this community of bacteria influence health and disease. One way to study the diversity of the microbiome is to take a sample, such as a section of stool, and perform DNA sequencing to determine which types of bacteria are present. This can reveal how the composition of the gut microbiome relates to our health, but cannot confirm whether these bacteria are the cause or the effect of most diseases. To overcome this problem, researchers need to be able to grow pure strains of these bacteria in order to unravel their underlying mechanisms. For over a century, the conventional way to cultivate bacteria has been to grow them in a Petri dish. However, this method promotes the growth of more abundant, fast-growing bacterial strains. This results in a huge disconnect between the bacteria grown in a Petri dish and the diversity within the human gut, which is hindering our understanding of gut health and disease. Now, Watterson et al. have built a machine that improves the speed and number of cultivated bacterial organisms, thus paving the way for more detailed investigations of the human gut microbiome. This new system works by growing bacteria in millions of miniscule droplets which can be physically separated to help the expansion of slower growing species. Watterson et al. cultivated bacterial cells from a stool sample from a single donor using the droplet system and compared this to traditional culturing methods. The droplet technology increased the number of different organisms that were able to grow by up to four times, including those that were rare or slow-growing. Bacteria in the donor stool were then screened for populations that were resistant to antibiotics. This identified 21 antibiotic resistant bacteria which only grew in the droplets and not in Petri dishes. This droplet-based technology will make it possible to study bacterial strains that were previously difficult to grow. Furthermore, this method could help identify whether stool from a donor contains any antibiotic resistant strains, which can lead to clinical complications once transplanted. In future, this new technology could be used in laboratories or hospitals to study the role of the gut microbiome in health and disease.

The Roles of an Aluminum Underlayer in the Biocompatibility and Mechanical Integrity of Vertically Aligned Carbon Nanotubes for Interfacing with Retinal Neurons.

Retinal implant devices are becoming an increasingly realizable way to improve the vision of patients blinded by photoreceptor degeneration. As an electrode material that can improve restored visual acuity, carbon nanotubes (CNTs) excel due to their nanoscale topography, flexibility, surface chemistry, and double-layer capacitance. If vertically aligned carbon nanotubes (VACNTs) are biocompatible with retinal neurons and mechanically robust, they can further improve visual acuity-most notably in subretinal implants-because they can be patterned into high-aspect-ratio, micrometer-size electrodes. We investigated the role of an aluminum (Al) underlayer beneath an iron (Fe) catalyst layer used in the growth of VACNTs by chemical vapor deposition (CVD). In particular, we cultured dissociated retinal cells for three days in vitro (DIV) on unfunctionalized and oxygen plasma functionalized VACNTs grown from a Fe catalyst (Fe and Fe + Pl preparations, where Pl signifies the plasma functionalization) and an Fe catalyst with an Al underlayer (Al/Fe and Al/Fe + Pl preparations). The addition of the Al layer increased the mechanical integrity of the VACNT interface and enhanced retinal neurite outgrowth over the Fe preparation. Unexpectedly, the extent of neurite outgrowth was significantly greater in the Al/Fe than in the Al/Fe+Pl preparation, suggesting plasma functionalization can negatively impact biocompatibility for some VACNT preparations. Additionally, we show our VACNT growth process for the Al/Fe preparation can support neurite outgrowth for up to 7 DIV. By demonstrating the retinal neuron biocompatibility, mechanical integrity, and pattern control of our VACNTs, this work offers VACNT electrodes as a solution for improving the restored visual acuity provided by modern retinal implants.

Fractal solar panels: Optimizing aesthetic and electrical performances.

Solar energy technologies have been plagued by their limited visual appeal. Because the electrical power generated by solar panels increases with their surface area and therefore their occupancy of the observer's visual field, aesthetics will play an increasingly critical role in their future success in urban environments. Inspired by previous psychology research highlighting the aesthetic qualities of fractal patterns, we investigated panel designs featuring fractal electrodes. We conducted behavioral studies which compared observers' preferences for fractal and conventional bus-bar electrode patterns, along with computer simulations which compared their electrical performances. This led us to develop a hybrid electrode pattern which best combines the fractal and bus-bar designs. Here we show that the new hybrid electrode matches the electrical performance of bus-bars in terms of light transmission and minimizing electrical power losses, while benefiting from the superior aesthetics of fractal patterns. This innovative integration of psychology and engineering studies provides a framework for developing novel electrode patterns with increased implementation and acceptance.

Modeling the Improved Visual Acuity Using Photodiode Based Retinal Implants Featuring Fractal Electrodes.

Electronically restoring vision to patients blinded by severe retinal degenerations is rapidly becoming a realizable feat through retinal implants. Upon receiving an implant, previously blind patients can now detect light, locate objects, and determine object motion direction. However, the restored visual acuity (VA) is still significantly below the legal blindness level (VA < 20/200). The goal of this research is to optimize the inner electrode geometry in photovoltaic subretinal implants in order to restore vision to a VA better than blindness level. We simulated neural stimulation by 20 µm subretinal photovoltaic implants featuring square or fractal inner electrodes by: (1) calculating the voltage generated on the inner electrode based on the amount of light entering the photodiode, (2) mapping how this voltage spreads throughout the extracellular space surrounding retinal bipolar neurons, and (3) determining if these extracellular voltages are sufficient for neural stimulation. By optimizing the fractal inner electrode geometry, we show that all neighboring neurons can be stimulated using an irradiance of 12 mW/mm2, while the optimized square only stimulates ~10% of these neurons at an equivalent irradiance. The 20 µm fractal electrode can thus theoretically restore VA up to 20/80, if other limiting factors common to retinal degenerations, such as glia scarring and rewiring of retinal circuits, could be reduced. For the optimized square to stimulate all neighboring neurons, the irradiance has to be increased by almost 300%, which is very near the maximum permissible exposure safety limit. This demonstration that fractal electrodes can stimulate targeted neurons for long periods using safe irradiance levels highlights the possibility for restoring vision to a VA better than the blindness level using photodiode-based retinal implants.