All-graphene-based open fluidics for pumpless, small-scale fluid transport via laser-controlled wettability patterning.

Open microfluidics have emerged as a low-cost, pumpless alternative strategy to conventional microfluidics for delivery of fluid for a wide variety of applications including rapid biochemical analysis and medical diagnosis. However, creating open microfluidics by tuning the wettability of surfaces typically requires sophisticated cleanroom processes that are unamenable to scalable manufacturing. Herein, we present a simple approach to develop open microfluidic platforms by manipulating the surface wettability of spin-coated graphene ink films on flexible polyethylene terephthalate via laser-controlled patterning. Wedge-shaped hydrophilic tracks surrounded by superhydrophobic walls are created within the graphene films by scribing micron-sized grooves into the graphene with a CO2 laser. This scribing process is used to make superhydrophobic walls (water contact angle ∼160°) that delineate hydrophilic tracks (created through an oxygen plasma pretreatment) on the graphene for fluid transport. These all-graphene open microfluidic tracks are capable of transporting liquid droplets with a velocity of 20 mm s-1 on a level surface and uphill at elevation angles of 7° as well as transporting fluid in bifurcating cross and tree branches. The all-graphene open microfluidic manufacturing technique is rapid and amenable to scalable manufacturing, and consequently offers an alternative pumpless strategy to conventional microfluidics and creates possibilities for diverse applications in fluid transport.

Fabrication of High-resolution Graphene-based Flexible Electronics via Polymer Casting.

In this study, a novel method based on the transfer of graphene patterns from a rigid or flexible substrate onto a polymeric film surface via solvent casting was developed. The method involves the creation of predetermined graphene patterns on the substrate, casting a polymer solution, and directly transferring the graphene patterns from the substrate to the surface of the target polymer film via a peeling-off method. The feature sizes of the graphene patterns on the final film can vary from a few micrometers (as low as 5 µm) to few millimeters range. This process, applied at room temperature, eliminates the need for harsh post-processing techniques and enables creation of conductive graphene circuits (sheet resistance: ~0.2 kΩ/sq) with high stability (stable after 100 bending and 24 h washing cycles) on various polymeric flexible substrates. Moreover, this approach allows precise control of the substrate properties such as composition, biodegradability, 3D microstructure, pore size, porosity and mechanical properties using different film formation techniques. This approach can also be used to fabricate flexible biointerfaces to control stem cell behavior, such as differentiation and alignment. Overall, this promising approach provides a facile and low-cost method for the fabrication of flexible and stretchable electronic circuits.

Stamped multilayer graphene laminates for disposable in-field electrodes: application to electrochemical sensing of hydrogen peroxide and glucose.

A multi-step approach is described for the fabrication of multi-layer graphene-based electrodes without the need for ink binders or post-print annealing. Graphite and nanoplatelet graphene were chemically exfoliated using a modified Hummers' method and the dried material was thermally expanded. Expanded materials were used in a 3D printed mold and stamp to create laminate electrodes on various substrates. The laminates were examined for potential sensing applications using model systems of peroxide (H2O2) and enzymatic glucose detection. Within the context of these two assay systems, platinum nanoparticle electrodeposition and oxygen plasma treatment were examined as methods for improving sensitivity. Electrodes made from both materials displayed excellent H2O2 sensing capability compared to screen-printed carbon electrodes. Laminates made from expanded graphite and treated with platinum, detected H2O2 at a working potential of 0.3 V (vs. Ag/AgCl [0.1 M KCl]) with a 1.91 µM detection limit and sensitivity of 64 nA·µM-1·cm-2. Electrodes made from platinum treated nanoplatelet graphene had a H2O2 detection limit of 1.98 µM (at 0.3 V), and a sensitivity of 16.5 nA·µM-1·cm-2. Both types of laminate electrodes were also tested as glucose sensors via immobilization of the enzyme glucose oxidase. The expanded nanographene material exhibited a wide analytical range for glucose (3.7 µM to 9.9 mM) and a detection limit of 1.2 µM. The sensing range of laminates made from expanded graphite was slightly reduced (9.8 µM to 9.9 mM) and the detection limit for glucose was higher (18.5 µM). When tested on flexible substrates, the expanded graphite laminates demonstrated excellent adhesion and durability during testing. These properties make the electrodes adaptable to a variety of tests for field-based or wearable sensing applications. Graphical abstract Expanded graphite (eGR) and expanded nanoplatelet graphene (nGN) were chemically exfoliated, thermally expanded, and manually stamped into flexible multi-layer graphene laminate electrodes. Hydrogen peroxide amperometric testing of eGR laminates compared to nGN laminates and a screen printed carbon (SPC) electrode.

Printed Graphene Electrochemical Biosensors Fabricated by Inkjet Maskless Lithography for Rapid and Sensitive Detection of Organophosphates.

Solution phase printing of graphene-based electrodes has recently become an attractive low-cost, scalable manufacturing technique to create in-field electrochemical biosensors. Here, we report a graphene-based electrode developed via inkjet maskless lithography (IML) for the direct and rapid monitoring of triple-O linked phosphonate organophosphates (OPs); these constitute the active compounds found in chemical warfare agents and pesticides that exhibit acute toxicity as well as long-term pollution to soils and waterways. The IML-printed graphene electrode is nano/microstructured with a 1000 mW benchtop laser engraver and electrochemically deposited platinum nanoparticles (dia. ∼25 nm) to improve its electrical conductivity (sheet resistance decreased from ∼10 000 to 100 Ω/sq), surface area, and electroactive nature for subsequent enzyme functionalization and biosensing. The enzyme phosphotriesterase (PTE) was conjugated to the electrode surface via glutaraldehyde cross-linking. The resulting biosensor was able to rapidly measure (5 s response time) the insecticide paraoxon (a model OP) with a low detection limit (3 nM), and high sensitivity (370 nA/µM) with negligible interference from similar nerve agents. Moreover, the biosensor exhibited high reusability (average of 0.3% decrease in sensitivity per sensing event), stability (90% anodic current signal retention over 1000 s), longevity (70% retained sensitivity after 8 weeks), and the ability to selectively sense OP in actual soil and water samples. Hence, this work presents a scalable printed graphene manufacturing technique that can be used to create OP biosensors that are suitable for in-field applications as well as, more generally, for low-cost biosensor test strips that could be incorporated into wearable or disposable sensing paradigms.

High-Resolution Graphene Films for Electrochemical Sensing via Inkjet Maskless Lithography.

Solution-phase printing of nanomaterial-based graphene inks are rapidly gaining interest for fabrication of flexible electronics. However, scalable manufacturing techniques for high-resolution printed graphene circuits are still lacking. Here, we report a patterning technique [i.e., inkjet maskless lithography (IML)] to form high-resolution, flexible, graphene films (line widths down to 20 µm) that significantly exceed the current inkjet printing resolution of graphene (line widths ∼60 µm). IML uses an inkjet printed polymer lacquer as a sacrificial pattern, viscous spin-coated graphene, and a subsequent graphene lift-off to pattern films without the need for prefabricated stencils, templates, or cleanroom technology (e.g., photolithography). Laser annealing is employed to increase conductivity on thermally sensitive, flexible substrates [polyethylene terephthalate (PET)]. Laser annealing and subsequent platinum nanoparticle deposition substantially increases the electroactive nature of graphene as illustrated by electrochemical hydrogen peroxide (H2O2) sensing [rapid response (5 s), broad linear sensing range (0.1-550 µm), high sensitivity (0.21 µM/µA), and low detection limit (0.21 µM)]. Moreover, high-resolution, complex graphene circuits [i.e., interdigitated electrodes (IDE) with varying finger width and spacing] were created with IML and characterized via potassium chloride (KCl) electrochemical impedance spectroscopy (EIS). Results indicated that sensitivity directly correlates to electrode feature size as the IDE with the smallest finger width and spacing (50 and 50 µm) displayed the largest response to changes in KCl concentration (∼21 kΩ). These results indicate that the developed IML patterning technique is well-suited for rapid, solution-phase graphene film prototyping on flexible substrates for numerous applications including electrochemical sensing.

Enhanced enzymatic activity from phosphotriesterase trimer gold nanoparticle bioconjugates for pesticide detection.

The rapid detection of organophosphates (OPs), a class of strong neurotoxins, is critically important for monitoring acute insecticide exposure and potential chemical warfare agent use. Herein, we improve the enzymatic activity of a phosphotriesterase trimer (PTE3), an enzyme that selectively recognizes OPs directly, by conjugation with distinctly sized (i.e., 5, 10, and 20 nm diameter) gold nanoparticles (AuNPs). The number of enzymes immobilized on the AuNP was controlled by conjugating increasing molar ratios of PTE3 onto the AuNP surface via metal affinity coordination. This occurs between the PTE3-His6 termini and the AuNP-displayed Ni2+-nitrilotriacetic acid end groups and was confirmed with gel electrophoresis. The enzymatic efficiency of the resultant PTE3-AuNP bioconjugates was analyzed via enzyme progress curves acquired from two distinct assay formats that compared free unbound PTE3 with the following PTE3-AuNP bioconjugates: (1) fixed concentration of AuNPs while increasing the bioconjugate molar ratio of PTE3 displayed around the AuNP and (2) fixed concentration of PTE3 while increasing the bioconjugate molar ratio of PTE3-AuNP by decreasing the AuNP concentration. Both assay formats monitored the absorbance of p-nitrophenol that was produced as PTE3 hydrolyzed the substrate paraoxon, a commercial insecticide and OP nerve agent simulant. Results demonstrate a general equivalent trend between the two formats. For all experiments, a maximum enzymatic velocity (Vmax) increased by 17-fold over free enzyme for the lowest PTE3-AuNP ratio and the largest AuNP (i.e., ratio of 1 : 1, 20 nm dia. AuNP). This work provides a route to improve enzymatic OP detection strategies with enzyme-NP bioconjugates.

Electrical Differentiation of Mesenchymal Stem Cells into Schwann-Cell-Like Phenotypes Using Inkjet-Printed Graphene Circuits.

Graphene-based materials (GBMs) have displayed tremendous promise for use as neurointerfacial substrates as they enable favorable adhesion, growth, proliferation, spreading, and migration of immobilized cells. This study reports the first case of the differentiation of mesenchymal stem cells (MSCs) into Schwann cell (SC)-like phenotypes through the application of electrical stimuli from a graphene-based electrode. Electrical differentiation of MSCs into SC-like phenotypes is carried out on a flexible, inkjet-printed graphene interdigitated electrode (IDE) circuit that is made highly conductive (sheet resistance < 1 kΩ/sq) via a postprint pulse-laser annealing process. MSCs immobilized on the graphene printed IDEs and electrically stimulated/treated (etMSCs) display significant enhanced cellular differentiation and paracrine activity above conventional chemical treatment strategies [≈85% of the etMSCs differentiated into SC-like phenotypes with ≈80 ng mL-1 of nerve growth factor (NGF) secretion vs. 75% and ≈55 ng mL-1 for chemically treated MSCs (ctMSCs)]. These results help pave the way for in vivo peripheral nerve regeneration where the flexible graphene electrodes could conform to the injury site and provide intimate electrical simulation for nerve cell regrowth.

Enabling Inkjet Printed Graphene for Ion Selective Electrodes with Postprint Thermal Annealing.

Inkjet printed graphene (IPG) has recently shown tremendous promise in reducing the cost and complexity of graphene circuit fabrication. Herein we demonstrate, for the first time, the fabrication of an ion selective electrode (ISE) with IPG. A thermal annealing process in a nitrogen ambient environment converts the IPG into a highly conductive electrode (sheet resistance changes from 52.8 ± 7.4 MΩ/□ for unannealed graphene to 172.7 ± 33.3 Ω/□ for graphene annealed at 950 °C). Raman spectroscopy and field emission scanning electron microscopy (FESEM) analysis reveals that the printed graphene flakes begin to smooth at an annealing temperature of 500 °C and then become more porous and more electrically conductive when annealed at temperatures of 650 °C and above. The resultant thermally annealed, IPG electrodes are converted into potassium ISEs via functionalization with a poly(vinyl chloride) (PVC) membrane and valinomycin ionophore. The developed potassium ISE displays a wide linear sensing range (0.01-100 mM), a low detection limit (7 µM), minimal drift (8.6 × 10-6 V/s), and a negligible interference during electrochemical potassium sensing against the backdrop of interfering ions [i.e., sodium (Na), magnesium (Mg), and calcium (Ca)] and artificial eccrine perspiration. Thus, the IPG ISE shows potential for potassium detection in a wide variety of human fluids including plasma, serum, and sweat.

3D nanostructured inkjet printed graphene via UV-pulsed laser irradiation enables paper-based electronics and electrochemical devices.

Emerging research on printed and flexible graphene-based electronics is beginning to show tremendous promise for a wide variety of fields including wearable sensors and thin film transistors. However, post-print annealing/reduction processes that are necessary to increase the electrical conductivity of the printed graphene degrade sensitive substrates (e.g., paper) and are whole substrate processes that are unable to selectively anneal/reduce only the printed graphene-leaving sensitive device components exposed to damaging heat or chemicals. Herein a pulsed laser process is introduced that can selectively irradiate inkjet printed reduced graphene oxide (RGO) and subsequently improve the electrical conductivity (Rsheet∼0.7 kΩ□(-1)) of printed graphene above previously published reports. Furthermore, the laser process is capable of developing 3D petal-like graphene nanostructures from 2D planar printed graphene. These visible morphological changes display favorable electrochemical sensing characteristics-ferricyanide cyclic voltammetry with a redox peak separation (ΔEp) ≈ 0.7 V as well as hydrogen peroxide (H2O2) amperometry with a sensitivity of 3.32 µA mM(-1) and a response time of <5 s. Thus this work paves the way for not only paper-based electronics with graphene circuits, it enables the creation of low-cost and disposable graphene-based electrochemical electrodes for myriad applications including sensors, biosensors, fuel cells, and theranostic devices.