Development of a novel microsensor for the study of oxygen profiles in biofilms

Understanding of the processes taking place inside biofilms is a key parameter to progress in the optimization of biofiltration technologies. This study was conducted with the aim of developing a novel dissolved oxygen (DO) microsensor specially designed for biofilms monitoring. The microsensor was fabricated through standard photolithography techniques, resulting in a microelectrodes array (MEA) of 11 gold circular working electrodes, with a diameter of 50 µm , and a gold reference electrode, which allows obtaining a snapshot oxygen profile of 1 mm of depth. The performance of the sensor was fully characterized under different conditions, inwhich the sensor presented high sensitivity and repeatability, and low detection and quantification limits. Monitoring of sensor performance showed a stable and reliable response. The developed sensor was used in obtaining micropofiles in an aerobic heterotrophic biofilm, showing similar response to Clark-type commercial microsensors. These studies concluded that the novel MEA sensor for DO monitoring allows obtaining oxygen profiles within biofilms, becoming a useful tool for the research of many biological applications.Postprint (author's final draft

Bidirectional Modulation of Neuronal Cells Electrical and Mechanical Properties Through Pristine and Functionalized Graphene Substrates

[Abstract] In recent years, the quest for surface modifications to promote neuronal cell interfacing and modulation has risen. This course is justified by the requirements of emerging technological and medical approaches attempting to effectively interact with central nervous system cells, as in the case of brain-machine interfaces or neuroprosthetic. In that regard, the remarkable cytocompatibility and ease of chemical functionalization characterizing surface-immobilized graphene-based nanomaterials (GBNs) make them increasingly appealing for these purposes. Here, we compared the (morpho)mechanical and functional adaptation of rat primary hippocampal neurons when interfaced with surfaces covered with pristine single-layer graphene (pSLG) and phenylacetic acid-functionalized single-layer graphene (fSLG). Our results confirmed the intrinsic ability of glass-supported single-layer graphene to boost neuronal activity highlighting, conversely, the downturn inducible by the surface insertion of phenylacetic acid moieties. fSLG-interfaced neurons showed a significant reduction in spontaneous postsynaptic currents (PSCs), coupled to reduced cell stiffness and altered focal adhesion organization compared to control samples. Overall, we have here demonstrated that graphene substrates, both pristine and functionalized, could be alternatively used to intrinsically promote or depress neuronal activity in primary hippocampal cultures.This work was funded by the European Union’s Horizon 2020 Research and Innovation Program under the Grant Agreements 785219 and 881603 of the Graphene Flagship. DS acknowledges the support of the European Union’s Horizon 2020 Research and Innovation Program under the Marie Skłodowska-Curie grant agreement no. 838902. MP as the recipient of the AXA Bionanotechnology Chair, is grateful to the AXA Research Fund for financial support. This work was performed under the Maria de Maeztu Units of Excellence Program from the Spanish State Research Agency-grant no. MDM-2017- 0720. AC thanks Xunta de Galicia for his research grant Atracción de Talento (no. ED431H 2020/17). GR acknowledges funding from RYC-2016-21412. HH acknowledges funding from Juan de la Cierva – Incorporación no. IJC-2018-037396-IXunta de Galicia; ED431H 2020/1

Biofilm oxygen profiling using an array of microelectrodes on a microfabricated needle

A novel microelectrode array (DO-MEA) sensor was designed and fabricated using microelectromechanical systems technology on a needle for real time measurement of dissolved oxygen (DO). The sensor consisted of eleven gold disk microelectrodes and a rectangular auxiliary electrode along them. The sensor can also be operated with an integrated reference system. Three different sensor designs were fabricated, and their responses were fully characterized and evaluated. The DO-MEA sensor presented a linear response in the 0-8 mg DO·L-1 concentration range in water, displaying high sensitivity and repeatability. Knowledge of bacterial activity inside biofilms is key to the optimization of applied biotechnologies. The developed sensor was validated against a commercial Clark-type microelectrode overcoming its drawbacks, by profiling a heterotrophic biofilm cultivated in a flat-plate bioreactor. The DO-MEA sensor provided a multipoint, simultaneous dissolved oxygen snapshot profile inside a biofilm with high spatial resolution due to its micrometric dimensions, thus becoming a powerful tool for the research of many similar biological-based processes and applications.Peer ReviewedPostprint (author’s final draft

Versatile Graphene-Based Platform for Robust Nanobiohybrid Interfaces

Technologically useful and robust graphene-based interfaces for devices require the introduction of highly selective, stable, and covalently bonded functionalities on the graphene surface, whilst essentially retaining the electronic properties of the pristine layer. This work demonstrates that highly controlled, ultrahigh vacuum covalent chemical functionalization of graphene sheets with a thiol-terminated molecule provides a robust and tunable platform for the development of hybrid nanostructures in different environments. We employ this facile strategy to covalently couple two representative systems of broad interest: metal nanoparticles, via S-metal bonds, and thiol-modified DNA aptamers, via disulfide bridges. Both systems, which have been characterized by a multi-technique approach, remain firmly anchored to the graphene surface even after several washing cycles. Atomic force microscopy images demonstrate that the conjugated aptamer retains the functionality required to recognize a target protein. This methodology opens a new route to the integration of high-quality graphene layers into diverse technological platforms, including plasmonics, optoelectronics, or biosensing. With respect to the latter, the viability of a thiol-functionalized chemical vapor deposition graphene-based solution-gated field-effect transistor array was assessed

Concurrent functional ultrasound imaging with graphene-based DC-coupled electrophysiology as a platform to study slow brain signals and cerebral blood flow under control and pathophysiological brain states

Current methodology used to investigate how shifts in brain states associated with regional cerebral blood volume (CBV) change in deep brain areas, are limited by either the spatiotemporal resolution of the CBV techniques, and/or compatibility with electrophysiological recordings; particularly in relation to spontaneous brain activity and the study of individual events. Additionally, infraslow brain signals (&lt;0.1 Hz), including spreading depolarisations, DC-shifts and infraslow oscillations (ISO), are poorly captured by traditional AC-coupled electrographic recordings; yet these very slow brain signals can profoundly change CBV. To gain an improved understanding of how infraslow brain signals couple to CBV we present a new method for concurrent CBV with wide bandwidth electrophysiological mapping using simultaneous functional ultrasound imaging (fUS) and graphene-based field effect transistor (gFET) DC-coupled electrophysiological acquisitions. To validate the feasibility of this methodology visually-evoked neurovascular coupling (NVC) responses were examined. gFET recordings are not affected by concurrent fUS imaging, and epidural placement of gFET arrays within the imaging window did not deteriorate fUS signal quality. To examine directly the impact of infra-slow potential shifts on CBV, cortical spreading depolarisations (CSDs) were induced. A biphasic pattern of decreased, followed by increased CBV, propagating throughout the ipsilateral cortex, and a delayed decrease in deeper subcortical brain regions was observed. In a model of acute seizures, CBV oscillations were observed prior to seizure initiation. Individual seizures occurred on the rising phase of both infraslow brain signal and CBV oscillations. When seizures co-occurred with CSDs, CBV responses were larger in amplitude, with delayed CBV decreases in subcortical structures. Overall, our data demonstrate that gFETs are highly compatible with fUS and allow concurrent examination of wide bandwidth electrophysiology and CBV. This graphene-enabled technological advance has the potential to improve our understanding of how infraslow brain signals relate to CBV changes in control and pathological brain states.</p

Improved metal-graphene contacts for low-noise, high-density microtransistor arrays for neural sensing

Poor metal contact interfaces are one of the main limitations preventing unhampered access to the full potential of two-dimensional materials in electronics. Here we present graphene solution-gated field-effect-transistors (gSGFETs) with strongly improved linearity, homogeneity and sensitivity for small sensor sizes, resulting from ultraviolet ozone (UVO) contact treatment. The contribution of channel and contact region to the total device conductivity and flicker noise is explored experimentally and explained with a theoretical model. Finally, in-vitro recordings of flexible microelectrocorticography (μ-ECoG) probes were performed to validate the superior sensitivity of the UVO-treated gSGFET to brain-like activity. These results connote an important step towards the fabrication of high-density gSGFET μ-ECoG arrays with state-of-the-art sensitivity and homogeneity, thus demonstrating the potential of this technology as a versatile platform for the new generation of neural interfaces

Flexible Graphene Solution-Gated Field-Effect Transistors : Efficient Transducers for Micro-Electrocorticography

Brain-computer interfaces and neural prostheses based on the detection of electrocorticography (ECoG) signals are rapidly growing fields of research. Several technologies are currently competing to be the first to reach the market; however, none of them fulfill yet all the requirements of the ideal interface with neurons. Thanks to its biocompatibility, low dimensionality, mechanical flexibility, and electronic properties, graphene is one of the most promising material candidates for neural interfacing. After discussing the operation of graphene solution-gated field-effect transistors (SGFET) and characterizing their performance in saline solution, it is reported here that this technology is suitable for μ-ECoG recordings through studies of spontaneous slow-wave activity, sensory-evoked responses on the visual and auditory cortices, and synchronous activity in a rat model of epilepsy. An in-depth comparison of the signal-to-noise ratio of graphene SGFETs with that of platinum black electrodes confirms that graphene SGFET technology is approaching the performance of state-of-the art neural technologies

Flexible Graphene Solution-Gated Field-Effect Transistors : Efficient Transducers for Micro-Electrocorticography

Brain-computer interfaces and neural prostheses based on the detection of electrocorticography (ECoG) signals are rapidly growing fields of research. Several technologies are currently competing to be the first to reach the market; however, none of them fulfill yet all the requirements of the ideal interface with neurons. Thanks to its biocompatibility, low dimensionality, mechanical flexibility, and electronic properties, graphene is one of the most promising material candidates for neural interfacing. After discussing the operation of graphene solution-gated field-effect transistors (SGFET) and characterizing their performance in saline solution, it is reported here that this technology is suitable for μ-ECoG recordings through studies of spontaneous slow-wave activity, sensory-evoked responses on the visual and auditory cortices, and synchronous activity in a rat model of epilepsy. An in-depth comparison of the signal-to-noise ratio of graphene SGFETs with that of platinum black electrodes confirms that graphene SGFET technology is approaching the performance of state-of-the art neural technologies

Method for exfoliating and transferring graphene from a doped silicon carbide substrate to another substrate

La presente invención se refiere a un procedimiento de exfoliación y transferencia de grafeno de un sustrato de carburo de silicio dopado a otro sustrato, donde dicho procedimiento se basa en su exfoliación inducida por el burbujeo de hidrógeno que se produce en la electrolisis del agua. [ES]The present invention relates to a method for exfoliating and transferring graphene from a doped silicon carbide substrate to another substrate, the method being based on exfoliation induced by hydrogen bubbles produced in the electrolysis of water. [EN]Peer reviewedConsejo Superior de Investigaciones Científicas (España), Centro de Investigación Biomédica en Red (CIBER)A1 Solicitud de patente con informe sobre el estado de la técnic

Method for exfoliating and transferring graphene from a doped silicon carbide substrate to another substrate

The present invention refers to a method for exfoliating and transferring graphene from a doped silicon carbide substrate to another substrate, the method being based on exfoliation induced by hydrogen bubbles produced in the electrolysis of waterPeer reviewedConsejo Superior de Investigaciones Científicas (España), Centro de Investigación Biomédica en Red (CIBER)A1 Solicitud de patente con informe sobre el estado de la técnic