Laser-induced graphene trending in biosensors: understanding electrode shelf-life of this highly porous material.

Laser-induced graphene (LIG) has received much attention in recent years as a possible transducer material for electroanalytical sensors. Its simplicity of fabrication and good electrochemical performance are typically highlighted. However, we found that unmodified and untreated LIG electrodes had a limited shelf-life for certain electroanalytical applications, likely due to the adsorption of adventitious hydrocarbons from the storage environment. Electrode responses did not change immediately after exposure to ambient conditions but over longer periods of time, probably due to the immense specific surface area of the LIG material. LIG shelf-life is seldomly discussed prominently in the literature, yet overall trends for solutions to this challenge can be identified. Such findings from the literature regarding the long-term storage stability of LIG electrodes, pure and modified, are discussed here along with explanations for likely protective mechanisms. Specifically, applying a protective coating on LIG electrodes after manufacture is possibly the easiest method to preserve electrode functionality and should be identified as a trend for well-performing LIG electrodes in the future. Furthermore, suggested influences of the accompanying LIG microstructure/morphology on electrode characteristics are evaluated.

Polypyrrole-palladium nanocomposite as a high-efficiency transducer for thrombin detection with liposomes as a label.

Sensitive and selective determination of protein biomarkers with high accuracy often remains a great challenge due to their existence in the human body at an exceptionally low concentration level. Therefore, sensing mechanisms that are easy to use, simple, and capable of accurate quantification of analyte are still in development to detect biomarkers at a low concentration level. To meet this end, we demonstrated a methodology to detect thrombin in serum at low concentration levels using polypyrrole (PPy)-palladium (Pd)nanoparticle-based hybrid transducers using liposomes encapsulated redox marker as a label. The morphology of Ppy-Pd composites was characterized by scanning electron microscopy, and the hybrid structure provided excellent binding and detection platform for thrombin detection in both buffer and serum solutions. For quantitative measurement of thrombin in PBS and serum, the change in current was monitored using differential pulse voltammetry, and the calculated limit of quantification (LOQ) and limit of detection (LOD) for the linear segment (0.1-1000 nM of thrombin) were 1.1 pM and 0.3 pM, in serum, respectively. The sensors also exhibited good stability and excellent selectivity towards the detection of thrombin, and thus make it a strong candidate for adopting its sensing applications in biomarker detection technologies.

Process-property correlations in laser-induced graphene electrodes for electrochemical sensing.

Laser-induced graphene (LIG) has emerged as a promising electrode material for electrochemical point-of-care diagnostics. LIG offers a large specific surface area and excellent electron transfer at low-cost in a binder-free and rapid fabrication process that lends itself well to mass production outside of the cleanroom. Various LIG micromorphologies can be generated when altering the energy input parameters, and it was investigated here which impact this has on their electroanalytical characteristics and performance. Energy input is well controlled by the laser power, scribing speed, and laser pulse density. Once the threshold of required energy input is reached a broad spectrum of conditions leads to LIG with micromorphologies ranging from delicate irregular brush structures obtained at fast, high energy input, to smoother and more wall like albeit still porous materials. Only a fraction of these LIG structures provided high conductance which is required for appropriate electroanalytical performance. Here, it was found that low, frequent energy input provided the best electroanalytical material, i.e., low levels of power and speed in combination with high spatial pulse density. For example, the sensitivity for the reduction of K3[Fe(CN)6] was increased almost 2-fold by changing fabrication parameters from 60% power and 100% speed to 1% power and 10% speed. These general findings can be translated to any LIG fabrication process independent of devices used. The simple fabrication process of LIG electrodes, their good electroanalytical performance as demonstrated here with a variety of (bio)analytically relevant molecules including ascorbic acid, dopamine, uric acid, p-nitrophenol, and paracetamol, and possible application to biological samples make them ideal and inexpensive transducers for electrochemical (bio)sensors, with the potential to replace the screen-printed systems currently dominating in on-site sensors used.

Laser-induced graphene interdigitated electrodes for label-free or nanolabel-enhanced highly sensitive capacitive aptamer-based biosensors.

Highly porous laser-induced graphene (LIG) is easily generated in complex electrode configurations such as interdigitated electrodes (IDEs). Here, we demonstrate that their superior capacitive response at low frequencies can be exploited in affinity biosensors using thrombin aptamers as model biorecognition elements. Of specific interest was the effect of electrode surface area on capacitance detection, and the comparison between a label-free format and enhancement strategies afforded by carboxy group bearing polymeric nanoparticles or liposomes. Electrochemical impedance spectroscopy (EIS) was used to investigate the LIG performance and optimize the biosensor design. Interestingly, the label-free strategy performed extremely well and additional labels decreased the limit of detection or increased the sensitivity only minimally. It is assumed that the highly porous nature of the LIG structures dominates the capacitive response so that labels removed from the surface have only limited influence Also, while slight performance changes can be observed for smaller vs. larger electrode structures, the performance of a LIG IDE is reasonably independent of its size. In the end, a dynamic range of 5 orders of magnitude was obtained (0.01 nM-1000 nM) with a limit of detection as low as 0.12 pM. When measured in serum, this increased to 1.3 pM. The good reproducibility (relative standard deviation (RSD), 4.90%) and repeatability (RSD, 2.59%) and good long-term stability (>7 weeks at 4 °C) prove that a LIG-based capacitance sensor is an excellent choice for affinity-based biosensor. The ease-of-production, the simplicity of modification and the superior performance even in a label-free format indicate that LIG-based biosensors should be considered in point-of-care diagnostics in the future.

A Robust strategy enabling addressable porous 3D carbon-based functional nanomaterials in miniaturized systems.

3D-porous carbon nanomaterials and their hybrids are ideal materials for energy storage and conversion, biomedical research, and wearable sensors, yet today's fabrication methods are too complicated and inefficient to implement into miniaturized systems. Instead, it is shown here that 3D-carbon nanofibrous electrodes of various designs, shapes and sizes, on flexible substrates, under ambient conditions and without complicated equipment and procedures can simply be "written" via a one-step laser-induced carbonization on electrospun nanofibers. Analytical functionalities are realized as full control over native polymer chemistry doping of the polymer (e.g. with metals) is provided. Similarly, being able to control mat morphology and its impact on the electroanalytical performance was studied. Ultimately, optimized writing conditions were harnessed for superior (bio)analytical sensing of important biomarkers (NADH, dopamine). The new procedure hence paves the way for future controlled studies on this 3D nanomaterial, for a multitude of functionalization and design possibilities, and for mass production capabilities necessary for their application in the real world.

Isolation and amplification of mRNA within a simple microfluidic lab on a chip.

The major modules for realizing molecular biological assays in a micro-total analysis system (µTAS) were developed for the detection of pathogenic organisms. The specific focus was the isolation and amplification of eukaryotic mRNA within a simple, single-channel device for very low RNA concentrations that could then be integrated with detection modules. The hsp70 mRNA from Cryptosporidium parvum was used as a model analyte. Important points of study were surface chemistries within poly(methyl methacrylate) (PMMA) microfluidic channels that enabled specific and sensitive mRNA isolation and amplification reactions for very low mRNA concentrations. Optimal conditions were achieved when the channel surface was carboxylated via UV/ozone treatment followed by the immobilization of polyamidoamine (PAMAM) dendrimers on the surface, thus increasing the immobilization efficiency of the thymidine oligonucleotide, oligo(dT)25, and providing a reliable surface for the amplification reaction, importantly, without the need for blocking agents. Additional chemical modifications of the remaining active surface groups were studied to avoid nonspecific capturing of nucleic acids and hindering of the mRNA amplification at low RNA concentrations. Amplification of the mRNA was accomplished using nucleic acid sequence-based amplification (NASBA), an isothermal, primer-dependent technique. Positive controls consisting of previously generated NASBA amplicons could be diluted 10(15) fold and still result in successful on-chip reamplification. Finally, the successful isolation and amplification of mRNA from as few as 30 C. parvum oocysts was demonstrated directly on-chip and compared to benchtop devices. This is the first proof of successful mRNA isolation and NASBA-based amplification of mRNA within a simple microfluidic device in relevant analytical volumes.