Organic mixed ionic‐electronic conductors have emerged as a key material for the development of bioelectronic devices due to their soft mechanical properties, biocompatibility, and high volumetric capacitance. In particular, PEDOT:PSS has become a choice material because it is highly conductive, easily processible, and commercially available. However, PEDOT:PSS is dispersible in water, leading to delamination of films when exposed to biological environments. For this reason, chemical cross–linking agents such as (3‐glycidyloxypropyl)trimethoxysilane (GOPS) are used to stabilize PEDOT:PSS films in water, but at the cost of decreased electrical performance. Here, it is shown that PEDOT:PSS thin films become water‐stable by simply baking at high temperatures (>150 °C) for a short time (≈ 2 min). It is shown that heat‐treated PEDOT:PSS films are as stable as their chemically‐cross–linked counterparts, with their performance maintained for >20 days both in vitro and in vivo. The heat‐treated films eliminate electrically insulating cross–linkers, resulting in a 3× increase in volumetric capacitance. Applying thermal energy using a focused femtosecond laser enables direct patterning of 3D PEDOT:PSS microstructures. The thermal treatment method is compatible with a wide range of substrates and is readily substituted into existing workflows for manufacturing devices, enabling its rapid adoption in the field of bioelectronics.

Minimally invasive electrodes of cellular scale that approach a bio-integrative level of neural recording could enable the development of scalable brain machine interfaces that stably interface with the same neural populations over long period of time.In this paper, we designed and created NeuroRoots, a bio-mimetic multi-channel implant sharing similar dimension (10µm wide, 1.5µm thick), mechanical flexibility and spatial distribution as axon bundles in the brain. A simple approach of delivery is reported based on the assembly and controllable immobilization of the electrode onto a 35µm microwire shuttle by using capillarity and surface-tension in aqueous solution. Once implanted into targeted regions of the brain, the microwire was retracted leaving NeuroRoots in the biological tissue with minimal surgical footprint and perturbation of existing neural architectures within the tissue. NeuroRoots was implanted using a platform compatible with commercially available electrophysiology rigs and with measurements of interests in behavioral experiments in adult rats freely moving into maze. We demonstrated that NeuroRoots electrodes reliably detected action potentials for at least 7 weeks and the signal amplitude and shape remained relatively constant during long-term implantation.This research represents a step forward in the direction of developing the next generation of seamless brain-machine interface to study and modulate the activities of specific sub-populations of neurons, and to develop therapies for a plethora of neurological diseases.

Schematics of OECT with oxygen sensitive polymer (left) and air-stable polymer (right), showing electrochemical reaction with oxygen and cells cytotoxicity.

Safe and long-term electrical stimulation of neurons requires charge injection without damaging the electrode and tissue. A common strategy to diminish adverse effects includes the modification of electrodes with materials that increases the charge injection capacity. Due to its high capacitance, the conducting polymer PEDOT:PSS is a promising coating material; however, the neural stimulation performance in terms of stability and safety remains largely unexplored. Here, PEDOT:PSS-coated platinum (Pt-PEDOT:PSS) microelectrodes are examined for neural stimulation and compared to bare platinum (Pt) electrodes. Microelectrodes in a bipolar configuration are used to deliver current-controlled, biphasic pulses with charge densities ranging from 64 to 255 μC cm−2. Stimulation for 2 h deteriorates bare Pt electrodes through corrosion, whereas the PEDOT:PSS coating prevents dissolution of Pt and shows no degradation. Acute stimulation of primary cortical cells cultured as neurospheres shows similar dependency on charge density for Pt and Pt-PEDOT:PSS electrodes with a threshold of 127 μC cm−2 and increased calcium response for higher charge densities. Continuous stimulation for 2 h results in higher levels of cell survival for Pt-PEDOT:PSS electrodes. Reduced cell survival on Pt electrodes is most profound for neurospheres in proximity of the electrodes. Extending the stimulation duration to 6 h increases cell death for both types of electrodes; however, neurospheres on Pt-PEDOT:PSS devices still show significant viability whereas stimulation is fatal for nearly all cells close to the Pt electrodes. This work demonstrates the protective properties of PEDOT:PSS that can be used as a promising approach to extend electrode lifetime and reduce cell damage for safe and long-term neural stimulation.

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed electric fields (PEFs) are promising for the improvement of treatment efficiency. However, they rely on rigid electrodes that cause acute and chronic damage, especially in soft tissues such as the brain. In this work, flexible electronics were used to deliver PEFs to tumors and the biological response was evaluated with fluorescent microscopy. Interdigitated gold electrodes on a thin, transparent parylene-C substrate were coated with the conducting polymer PEDOT:PSS, resulting in a conformable and biocompatible device. The effects of PEFs on tumors and their microenvironment were examined using various biological models. First, monolayers of glioblastoma cells were cultured on top of the electrodes to investigate phenomena in vitro. As an intermediate step, an in ovo model was developed where engineered tumor spheroids were grafted in the embryonic membrane of a quail. Due to the absence of an immune system, this led to highly vascularized tumors. At this early stage of development, embryos have no immune system, and tumors are not recognized as foreign bodies. Thus, they can develop fast while developing their own vessels from the existing embryo vascular system, which represents a valuable 3D cancer model. Finally, flexible electrode delivery of PEFs was evaluated in a complete organism with a functional immune system, using a syngenic, orthograft (intracranial) mouse model. Tumor spheroids were grafted into the brain of transgenic multi-fluorescent mice prior to the implantation of flexible organic electrode devices. A sealed cranial window enabled multiphoton imaging of the tumor and its microenvironment during treatment with PEFs over a period of several weeks.

Transparent microelectrode arrays enable simultaneous electrical recording and optical imaging of neuronal networks in the brain. Electrodes made of the conducting polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS) are transparent; however, device fabrication necessitates specific processes to avoid deterioration of the organic material. Here, we present an innovative fabrication scheme for a neural probe that consists of transparent PEDOT:PSS electrodes and demonstrate its compatibility with 2-photon microscopy. The electrodes show suitable impedance to record local field potentials from the cortex of mice and sufficient transparency to visualize GCaMP6f-expressing neurons underneath the PEDOT:PSS features. The results validate the performance of the neural probe, which paves the way to study the complex dynamics of in vivo neuronal activity with both a high spatial and temporal resolution to better understand the brain.

Objective: Monitoring of impedance changes during electroporation-based treatments can be used to study the biological response and provide feedback regarding treatment progression. However, seamless integration of the sensing electrodes with the setup can be challenging and high impedance sensing electrodes limit the recording sensitivity as well as the spatial resolution. Here, we present an all-in-one microchip containing stimulation electrodes, as well as an array of low impedance, micro-scale sensing electrodes for highly sensitive impedance monitoring. Methods: An in vitro platform is fabricated with integrated stimulation and sensing electrodes. To reduce the impedance, the sensing electrodes are coated with the conducting polymer PEDOT:PSS. The performance is studied during the growth of a confluent cell layer and treatment with electrical pulses. Results: Coated electrodes, compared to uncoated electrodes, show more pronounced impedance changes in a broader frequency range throughout the formation of a confluent cell layer and after electrical treatment. Conclusion: PEDOT:PSS coatings enhance the monitoring of impedance changes with micro-scale electrodes, enabling high spatial resolution and increased sensitivity. Significance: Such monitoring systems can be used to study electroporation dynamics and monitor treatment progression for better understanding of underlying mechanisms and improved outcomes.

Glioblastoma Multiforme is a highly lethal form of brain cancer, resistant to traditional therapeutic approaches and oftentimes hardly resectable. The application of pulsed electric fields (PEF) is gaining prominence as a highly effective approach for combating malignant tumors. However, PEF application at high voltages can generate reactive oxygen species through electrochemical events at electrodes, which can greatly affect intracellular processes and damage healthy cells. Here, we present an in depth study on the cellular impact of coating metal electrodes with an organic polymer PEDOT:PSS. We compared the effect of PEF application through coated and uncoated gold electrodes on the U87 human glioblastoma cell line. The results show that PEF application using PEDOT:PSS-coated electrodes does not induce intracellular ROS generation, even at high voltages, contrary to that observed with uncoated electrodes. PEF delivery with PEDOT:PSS coated electrodes results in minimal cell electroporation and a lower intracellular calcium response than uncoated metal electrodes. The application of the antioxidant MnTBAP allowed us to establish that superoxide generation is partially responsible for the higher intracellular calcium response observed in uncoated metal electrodes. The results demonstrate that PEDOT-coated electrodes allow for PEF application without intracellular ROS generation, with the trade-off being a diminished electroporation efficiency. These electrodes could therefore be useful for PEF application in ROS-sensitive tissues, as well as for disentangling the effect of PEFs on cells from the metabolic impact of electrolytic events arising from the electrode material.

Glioblastoma is a highly aggressive brain tumor, very invasive and thus difficult to eradicate with standard oncology therapies. Bioelectric treatments based on pulsed electric fields have proven to be a successful method to treat cancerous tissues. However, they rely on stiff electrodes, which cause acute and chronic injuries, especially in soft tissues like the brain. Here we demonstrate the feasibility of delivering pulsed electric fields with flexible electronics using an in ovo vascularized tumor model. We show with fluorescence widefield and multiphoton microscopy that pulsed electric fields induce vasoconstriction of blood vessels and evoke calcium signals in vascularized glioblastoma spheroids stably expressing a genetically encoded fluorescence reporter. Simulations of the electric field delivery are compared with the measured influence of electric field effects on cell membrane integrity in exposed tumor cells. Our results confirm the feasibility of flexible electronics as a means of delivering intense pulsed electric fields to tumors in an intravital 3D vascularized model of human glioblastoma.

The combination of Ca2+ ions and electroporation has gained attention as potential alternative to electrochemotherapy. Ca2+ is an important component of the cell membrane repair system and its presence directly influences the dynamics of the pore cycle after electroporation which can be exploited for cancer therapies. Here, the influence of Ca2+ concentration is investigated on small molecule electrotransfer and release of Calcein from 4T1, MX-1, B16F10, U87 cancer cells after cell exposure to microsecond electric pulses. Moreover, we investigated simultaneous molecule electrotransfer and intracellular calcium ion influx when media was supplemented with different Ca2+ concentrations. Results show that increased concentrations of calcium ions reduce the electrotransfer of small molecules to different lines of cancer cells as well as the release of Calcein. These effects are related with an enhanced membrane repair mechanism. Overall, we show that the efficiency of molecular electrotransfer can be controlled by regulating Ca2+ concentration in the electroporation medium. For the first time, the cause of cancer cell death in vitro from 1 mM CaCl2 concentrations is related to the irreversible loss of Ca2+ homeostasis after cell electroporation. Our findings provide fundamental insight on the mechanisms of Ca2+ electroporation that might lead to improved therapeutic outcomes.

Electroporation or electropermeabilization occurs when cells are exposed to sufficiently high-pulsed electric fields. This phenomenon is now an important technique to facilitate the transmembrane transport of molecules, however, the role of the electrode material and associated electrochemical events remain unclear. Here, it is shown that electrical stimulation with PEDOT:PSS-coated electrodes strongly reduces irreversible electrochemical reactions that otherwise lead to changes in pH and the generation of reactive oxygen species. Coated electrodes exhibit a slightly higher threshold for cell stimulation, however, little electropermeabilization is observed at voltages that greatly exceed the threshold for uncoated electrodes. The findings indicate that electrically induced, extracellular chemical changes represent an important mechanism in the cellular electropermeabilization response. Moreover, PEDOT:PSS-coated electrodes separate the delivery of electric pulses from chemical changes, which paves the way for future experiments that could lead to a better understanding of underlying mechanisms.

Poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS) is widely used as a coating on microelectrode arrays in order to reduce impedance for both in vitro and in vivo electrophysiology. In many applications, electrode performance of months to years is desired; yet, there are few studies to date that examine the long-term stability of conducting polymers and their devices. Here, the stability of PEDOT:PSS microelectrodes is examined over a period of four months in cell culture media enriched with fetal bovine serum. The electrochemical impedance remains constant for most electrodes throughout the study, and only small changes in the structure of functional electrodes are observed. The results demonstrate that PEDOT:PSS electrodes show adequate stability for a variety of in vitro electrophysiology applications in toxicology, drug development, tissue engineering, and fundamental studies of electrically active cells and tissues.

Poly(3,4‐ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS) is a conducting polymer commonly used to coat the electrodes of biomedical devices to reduce impedance and improve charge injection. The stimulation performance of such electrodes depends on coating thickness due to the volumetric properties of the conducting polymer. Here, the influence of PEDOT:PSS coating thickness on impedance, charge injection, and electrode stability is examined. The results show that stacking spin‐coated layers is a reliable method to control coating thickness without introducing electronic or ionic blocking elements in between layers. Besides improving the polymer‐electrolyte interaction due to a higher capacitance, thicker coatings can endure more stimulation pulses and withstand higher voltages without loss of performance, thus enhancing the lifetime of the electrode. This work presents fundamental insights into the coupling of PEDOT:PSS coated electrodes with electrolytes and provides guidance in the development of organic bioelectronic stimulation devices for optimum performance and longevity.

We demonstrate the ability to excite atoms at well-defined, programmable locations in a magnetooptical trap, either to the continuum (ionization), or to a Rydberg state. To this end, excitation laser light is shaped into arbitrary intensity patterns with a spatial light modulator. These optical patterns are sensitive to aberrations of the phase of the light field, occurring while traversing the optical beamline. These aberrations are characterized and corrected without observing the actual light field in the vacuum chamber.