High-Performance, Easy-to-Fabricate, Nanocomposite Heater for Life Sciences and Biomedical Applications.

Microheaters are used in several applications, including medical diagnostics, synthesis, environmental monitoring, and actuation. Conventional microheaters rely on thin-film electrodes microfabricated in a clean-room environment. However, low-cost alternatives based on conductive paste electrodes fabricated using printing techniques have started to emerge over the years. Here, we report a surprising effect that leads to significant electrode performance improvement as confirmed by the thorough characterization of bulk, processed, and conditioned samples. Mixing silver ink and PVA results in the solubilization of performance-hindering organic compounds. These compounds evaporate during heating cycles. The new electrodes, which reach a temperature of 80 °C within 5 min using a current of 7.0 A, display an overall 42% and 35% improvement in the mechanical (hardness) and electrical (resistivity) properties compared to pristine silver ink electrodes. To validate our results, we use the composite heater to amplify and detect parasite DNA from Trypanosoma brucei, associated with African sleeping sickness. Our LAMP test compares well with commercially available systems, confirming the excellent performance of our nanocomposite heaters. Since their fabrication relies on well-established techniques, we anticipate they will find use in a range of applications.

Review of Physical Layer Security in Molecular Internet of Nano-Things.

Molecular networking has been identified as a key enabling technology for Internet-of-Nano-Things (IoNT): microscopic devices that can monitor, process information, and take action in a wide range of medical applications. As the research matures into prototypes, the cybersecurity challenges of molecular networking are now being researched on at both the cryptographic and physical layer level. Due to the limited computation capabilities of IoNT devices, physical layer security (PLS) is of particular interest. As PLS leverages on channel physics and physical signal attributes, the fact that molecular signals differ significantly from radio frequency signals and propagation means new signal processing methods and hardware is needed. Here, we review new vectors of attack and new methods of PLS, focusing on 3 areas: (1) information theoretical secrecy bounds for molecular communications, (2) key-less steering and decentralized key-based PLS methods, and (3) new methods of achieving encoding and encryption through bio-molecular compounds. The review will also include prototype demonstrations from our own lab that will inform future research and related standardization efforts.

Effects of Processing-Induced Contamination on Organic Electronic Devices.

Organic semiconductors are a family of pi-conjugated compounds used in many applications, such as displays, bioelectronics, and thermoelectrics. However, their susceptibility to processing-induced contamination is not well understood. Here, it is shown that many organic electronic devices reported so far may have been unintentionally contaminated, thus affecting their performance, water uptake, and thin film properties. Nuclear magnetic resonance spectroscopy is used to detect and quantify contaminants originating from the glovebox atmosphere and common laboratory consumables used during device fabrication. Importantly, this in-depth understanding of the sources of contamination allows the establishment of clean fabrication protocols, and the fabrication of organic field effect transistors (OFETs) with improved performance and stability. This study highlights the role of unintentional contaminants in organic electronic devices, and demonstrates that certain stringent processing conditions need to be met to avoid scientific misinterpretation, ensure device reproducibility, and facilitate performance stability. The experimental procedures and conditions used herein are typical of those used by many groups in the field of solution-processed organic semiconductors. Therefore, the insights gained into the effects of contamination are likely to be broadly applicable to studies, not just of OFETs, but also of other devices based on these materials.

Platform-agnostic electrochemical sensing app and companion potentiostat.

Electrochemical sensing is ubiquitous in a number of fields ranging from biosensing, to environmental monitoring through to food safety and battery or corrosion characterisation. Whereas conventional potentiostats are ideal to develop assays in laboratory settings, they are in general, not well-suited for field work due to their size and power requirements. To address this need, a number of portable battery-operated potentiostats have been proposed over the years. However, most open source solutions do not take full advantage of integrated circuit (IC) potentiostats, a rapidly evolving field. This is partly due to the constraining requirements inherent to the development of dedicated interfaces, such as apps, to address and control a set of common electrochemical sensing parameters. Here we propose the PocketEC, a universal app that has all the functionalities to interface with potentiostat ICs through a user defined property file. The versatility of PocketEC, developed with an assay developer mindset, was demonstrated by interfacing it, via Bluetooth, to the ADuCM355 evaluation board, the open-source DStat potentiostat and the Voyager board, a custom-built, small footprint potentiostat based around the LMP91000 chip. The Voyager board is presented here for the first time. Data obtained using a standard redox probe, Ferrocene Carboxylic Acid (FCA) and a silver ion assay using anodic stripping multi-step amperometry were in good agreement with analogous measurements using a bench top potentiostat. Combined with its Voyager board companion, the PocketEC app can be used directly for a number of wearable or portable electrochemical sensing applications. Importantly, the versatility of the app makes it a candidate of choice for the development of future portable potentiostats. Finally, the app is available to download on the Google Play store and the source codes and design files for the PocketEC app and the Voyager board are shared via Creative Commons license (CC BY-NC 3.0) to promote the development of novel portable or wearable applications based on electrochemical sensing.

Flow Rate-Independent Multiscale Liquid Biopsy for Precision Oncology.

Immunoaffinity-based liquid biopsies of circulating tumor cells (CTCs) hold great promise for cancer management but typically suffer from low throughput, relative complexity, and postprocessing limitations. Here, we address these issues simultaneously by decoupling and independently optimizing the nano-, micro-, and macro-scales of an enrichment device that is simple to fabricate and operate. Unlike other affinity-based devices, our scalable mesh approach enables optimum capture conditions at any flow rate, as demonstrated with constant capture efficiencies, above 75% between 50 and 200 µL min-1. The device achieved 96% sensitivity and 100% specificity when used to detect CTCs in the blood of 79 cancer patients and 20 healthy controls. We demonstrate its postprocessing capacity with the identification of potential responders to immune checkpoint inhibition (ICI) therapy and the detection of HER2 positive breast cancer. The results compare well with other assays, including clinical standards. This suggests that our approach, which overcomes major limitations associated with affinity-based liquid biopsies, could help improve cancer management.

Multifunctional Composite Hydrogels for Bacterial Capture, Growth/Elimination, and Sensing Applications.

Hydrogels are cross-linked networks of hydrophilic polymer chains with a three-dimensional structure. Owing to their unique features, the application of hydrogels for bacterial/antibacterial studies and bacterial infection management has grown in importance in recent years. This trend is likely to continue due to the rise in bacterial infections and antimicrobial resistance. By exploiting their physicochemical characteristics and inherent nature, hydrogels have been developed to achieve bacterial capture and detection, bacterial growth or elimination, antibiotic delivery, or bacterial sensing. Traditionally, the development of hydrogels for bacterial/antibacterial studies has focused on achieving a single function such as antibiotic delivery, antibacterial activity, bacterial growth, or bacterial detection. However, recent studies demonstrate the fabrication of multifunctional hydrogels, where a single hydrogel is capable of performing more than one bacterial/antibacterial function, or composite hydrogels consisting of a number of single functionalized hydrogels, which exhibit bacterial/antibacterial function synergistically. In this review, we first highlight the hydrogel features critical for bacterial studies and infection management. Then, we specifically address unique hydrogel properties, their surface/network functionalization, and their mode of action for bacterial capture, adhesion/growth, antibacterial activity, and bacterial sensing, respectively. Finally, we provide insights into different strategies for developing multifunctional hydrogels and how such systems can help tackle, manage, and understand bacterial infections and antimicrobial resistance. We also note that the strategies highlighted in this review can be adapted to other cell types and are therefore likely to find applications beyond the field of microbiology.

Graphene FET Sensors for Alzheimer's Disease Protein Biomarker Clusterin Detection.

We report on the fabrication and characterisation of graphene field-effect transistor (GFET) biosensors for the detection of Clusterin, a prominent protein biomarker of Alzheimer's disease (AD). The GFET sensors were fabricated on Si/SiO2 substrate using photolithographic patterning and metal lift-off techniques with evaporated chromium and sputtered gold contacts. Raman Spectroscopy was performed on the devices to determine the quality of the graphene. The GFETs were annealed to improve their performance before the channels were functionalized by immobilising the graphene surface with linker molecules and anti-Clusterin antibodies. Concentration of linker molecules was also independently verified by absorption spectroscopy using the highly collimated micro-beam light of Diamond B23 beamline. The detection was achieved through the binding reaction between the antibody and varying concentrations of Clusterin antigen from 1 to 100 pg/mL, as well as specificity tests using human chorionic gonadotropin (hCG), a glycoprotein risk biomarker of certain cancers. The GFETs were characterized using direct current (DC) 4-probe electrical resistance (4-PER) measurements, which demonstrated a limit of detection of the biosensors to be ∼ 300 fg/mL (4 fM). Comparison with back-gated Dirac voltage shifts with varying concentration of Clusterin show 4-PER measurements to be more accurate, at present, and point to a requirement for further optimisation of the fabrication processes for our next generation of GFET sensors. Thus, we have successfully fabricated a promising set of GFET biosensors for the detection of Clusterin protein biomarker. The developed GFET biosensors are entirely generic and also have the potential to be applied to a variety of other disease detection applications such as Parkinson's, cancer, and cardiovascular.

Label-Free Protein Analysis Using Liquid Chromatography with Gravimetric Detection.

The detection and analysis of proteins in a label-free manner under native solution conditions is an increasingly important objective in analytical bioscience platform development. Common approaches to detect native proteins in solution often require specific labels to enhance sensitivity. Dry mass sensing approaches, by contrast, using mechanical resonators, can operate in a label-free manner and offer attractive sensitivity. However, such approaches typically suffer from a lack of analyte selectivity as the interface between standard protein separation techniques and micro-resonator platforms is often constrained by qualitative mechanical sensor performance in the liquid phase. Here, we describe a strategy that overcomes this limitation by coupling liquid chromatography with a quartz crystal microbalance (QCM) platform by using a microfluidic spray dryer. We explore a strategy which allows first to separate a protein mixture in a physiological buffer solution using size exclusion chromatography, permitting specific protein fractions to be selected, desalted, and subsequently spray-dried onto the QCM for absolute mass analysis. By establishing a continuous flow interface between the chromatography column and the spray device via a flow splitter, simultaneous protein mass detection and sample fractionation is achieved, with sensitivity down to a 100 µg/mL limit of detection. This approach for quantitative label-free protein mixture analysis offers the potential for detection of protein species under physiological conditions.

Low-Cost Microfabrication Tool Box.

Microsystems are key enabling technologies, with applications found in almost every industrial field, including in vitro diagnostic, energy harvesting, automotive, telecommunication, drug screening, etc. Microsystems, such as microsensors and actuators, are typically made up of components below 1000 microns in size that can be manufactured at low unit cost through mass-production. Yet, their development for commercial or educational purposes has typically been limited to specialized laboratories in upper-income countries due to the initial investment costs associated with the microfabrication equipment and processes. However, recent technological advances have enabled the development of low-cost microfabrication tools. In this paper, we describe a range of low-cost approaches and equipment (below £1000), developed or adapted and implemented in our laboratories. We describe processes including photolithography, micromilling, 3D printing, xurography and screen-printing used for the microfabrication of structural and functional materials. The processes that can be used to shape a range of materials with sub-millimetre feature sizes are demonstrated here in the context of lab-on-chips, but they can be adapted for other applications. We anticipate that this paper, which will enable researchers to build a low-cost microfabrication toolbox in a wide range of settings, will spark a new interest in microsystems.

Design and Development of a Disposable Lab-on-a-Chip for Prostate Cancer Detection.

We have designed and fabricated a low-cost modular electrical and fluidic integration platform for microelectromechanical systems (MEMS) based biosensors, paving the way for a disposable, low-cost Lab-on-a-Chip. We demonstrate seamless integration using an additive manufacturing enabled "plug-and-play" platform that does not require permanent electronic or fluidic integration. This paper describes the fabrication steps and assembly of the method and highlights its advantages over the more traditional methods, such as `wire bonding' and `flip chip'. We also provide design guidelines for improved biosensing, taking transport and binding kinetics into consideration in the context of prostate cancer diagnosis. Our novel approach combined with the design guidelines, opens up new opportunities for low-cost disposable high-density MEMS-based lab-on-a-chips for biosensing applications.

Resolving protein mixtures using microfluidic diffusional sizing combined with synchrotron radiation circular dichroism.

Circular dichroism spectroscopy has become a powerful tool to characterise proteins and other biomolecules. For heterogeneous samples such as those present for interacting proteins, typically only average spectroscopic features can be resolved. Here we overcome this limitation by using free-flow microfluidic size separation in-line with synchrotron radiation circular dichroism to resolve the secondary structure of each component of a model protein mixture containing monomers and fibrils. To enable this objective, we have integrated far-UV compatible measurement chambers into PDMS-based microfluidic devices. Two architectures are proposed so as to accommodate for a wide range of concentrations. The approach, which can be used in combination with other bulk measurement techniques, paves the way to the study of complex mixtures such as the ones associated with protein misfolding and aggregation diseases including Alzheimer's and Parkinson's diseases.

Microfluidic deposition for resolving single-molecule protein architecture and heterogeneity.

Scanning probe microscopy provides a unique window into the morphology, mechanics, and structure of proteins and their complexes on the nanoscale. Such measurements require, however, deposition of samples onto substrates. This process can affect conformations and assembly states of the molecular species under investigation and can bias the molecular populations observed in heterogeneous samples through differential adsorption. Here, we show that these limitations can be overcome with a single-step microfluidic spray deposition platform. This method transfers biological solutions to substrates as microdroplets with subpicoliter volume, drying in milliseconds, a timescale that is shorter than typical diffusion times of proteins on liquid-solid interfaces, thus avoiding surface mass transport and change to the assembly state. Finally, the single-step deposition ensures the attachment of the full molecular content of the sample to the substrate, allowing quantitative measurements of different molecular populations within heterogeneous systems, including protein aggregates.

Combining Affinity Selection and Specific Ion Mobility for Microchip Protein Sensing.

The sensitive detection of proteins is a key objective in many areas of biomolecular science, ranging from biophysics to diagnostics. However, sensing in complex biological fluids is hindered by nonspecific interactions with off-target species. Here, we describe and demonstrate an assay that utilizes both the chemical and physical properties of the target species to achieve high selectivity in a manner not possible by chemical complementarity alone, in complex media. We achieve this objective through a combinatorial strategy, by simultaneously exploiting free-flow electrophoresis for target selection, on the basis of electrophoretic mobility, and conventional affinity-based selection. In addition, we demonstrate amplification of the resultant signal by a catalytic DNA nanocircuit. This approach brings together the inherent solution-phase advantages of microfluidic sample handling with isothermal, enzyme-free signal amplification. With this method, no surface immobilization or washing steps are required, and our assay is well suited to monoepitopic targets, presenting advantages over conventional ELISA techniques.

Enhancing the Resolution of Micro Free Flow Electrophoresis through Spatially Controlled Sample Injection.

Free flow electrophoresis is a versatile technique for the continuous separation of mixtures with both preparative and analytical applications. Microscale versions of free flow electrophoresis are particularly attractive strategies because of their fast separation times, ability to work with small sample volumes, and large surface area to volume ratios facilitating rapid heat transfer, thus minimizing the detrimental effects of Joule heating even at high voltages. The resolution of microscale free flow electrophoresis, however, is limited by the broadening of the analyte beam in the microfluidic channel, an effect that becomes especially pronounced when the analyte is deflected significantly away from its original position. Here, we describe and demonstrate how restricting spatially the sample injection and collection to the regions where the gradients in the velocity distribution of the carrier medium are the smallest allows this broadening effect to be substantially suppressed and hence the resolution of microscale free flow electrophoresis devices to be increased. To demonstrate this concept, we fabricated microfluidic free flow electrophoresis devices with spatially restricted injection nozzles implemented through the use of multilayer soft-photolithography and further integrated quartz based observation areas for fluorescent detection and imaging. With these devices, we demonstrated a 5-fold reduction in the extent of beam broadening relative to conventional free flow electrophoresis approaches with nonrestricted sample introduction. The manifold enhancement in the achievable resolution of microscale free flow electrophoresis devices opens up the possibility of rapid separation and analysis of complex mixtures.

Biophotonics of Native Silk Fibrils.

Native silk fibroin (NSF) is a unique biomaterial with extraordinary mechanical and biochemical properties. These key characteristics are directly associated with the physical transformation of unstructured, soluble NSF into highly organized nano- and microscale fibrils rich in ß-sheet content. Here, it is shown that this NSF fibrillation process is accompanied by the development of intrinsic fluorescence in the visible range, upon near-UV excitation, a phenomenon that has not been investigated in detail to date. Here, the optical and fluorescence characteristics of NSF fibrils are probed and a route for potential applications in the field of self-assembled optically active biomaterials and systems is explored. In particular, it is demonstrated that NSF can be structured into autofluorescent microcapsules with a controllable level of ß-sheet content and fluorescence properties. Furthermore, a facile and efficient fabrication route that permits arbitrary patterns of NSF microcapsules to be deposited on substrates under ambient conditions is shown. The resulting fluorescent NSF patterns display a high level of photostability. These results demonstrate the potential of using native silk as a new class of biocompatible photonic material.

Microfluidics for Protein Biophysics.

Microfluidics has the potential to transform experimental approaches across the life sciences. In this review, we discuss recent advances enabled by the development and application of microfluidic approaches to protein biophysics. We focus on areas where key fundamental features of microfluidics open up new possibilities and present advantages beyond low volumes and short time-scale analysis, conventionally provided by microfluidics. We discuss the two most commonly used forms of microfluidic technology, single-phase laminar flow and multiphase microfluidics. We explore how the understanding and control of the characteristic physical features of the microfluidic regime, the integration of microfluidics with orthogonal systems and the generation of well-defined microenvironments can be used to develop novel devices and methods in protein biophysics for sample manipulation, functional and structural studies, detection and material processing.

Enhanced Quality Factor Label-free Biosensing with Micro-Cantilevers Integrated into Microfluidic Systems.

Microelectromechanical systems (MEMS) have enabled the development of a new generation of sensor platforms. Acoustic sensor operation in liquid, the native environment of biomolecules, causes, however, significant degradation of sensing performance due to viscous drag and relies on the availability of capture molecules to bind analytes of interest to the sensor surface. Here, we describe a strategy to interface MEMS sensors with microfluidic platforms through an aerosol spray. Our sensing platform comprises a microfluidic spray nozzle and a microcantilever array operated in dynamic mode within a closed loop oscillator. A solution containing the analyte is sprayed uniformly through picoliter droplets onto the microcantilever surface; the micrometer-scale drops evaporate rapidly and leave the solutes behind, adding to the mass of the cantilever. This sensing scheme results in a 50-fold increase in the quality factor compared to operation in liquid, yet allows the analytes to be introduced into the sensing system from a solution phase. It achieves a 370 femtogram limit of detection, and we demonstrate quantitative label-free analysis of inorganic salts and model proteins. These results demonstrate that the standard resolution limits of cantilever sensing in dynamic mode can be overcome with the integration of spray microfluidics with MEMS.

Microfluidic devices fabricated using fast wafer-scale LED-lithography patterning.

Current lithography approaches underpinning the fabrication of microfluidic devices rely on UV exposure of photoresists to define microstructures in these materials. Conventionally, this objective is achieved with gas discharge mercury lamps, which are capable of producing high intensity UV radiation. However, these sources are costly, have a comparatively short lifetime, necessitate regular calibration, and require significant time to warm up prior to exposure taking place. To address these limitations we exploit advances in solid state sources in the UV range and describe a fast and robust wafer-scale laboratory exposure system relying entirely on UV-Light emitting diode (UV-LED) illumination. As an illustration of the potential of this system for fast and low-cost microfluidic device production, we demonstrate the microfabrication of a 3D spray-drying microfluidic device and a 3D double junction microdroplet maker device.

High operational and environmental stability of high-mobility conjugated polymer field-effect transistors through the use of molecular additives.

Due to their low-temperature processing properties and inherent mechanical flexibility, conjugated polymer field-effect transistors (FETs) are promising candidates for enabling flexible electronic circuits and displays. Much progress has been made on materials performance; however, there remain significant concerns about operational and environmental stability, particularly in the context of applications that require a very high level of threshold voltage stability, such as active-matrix addressing of organic light-emitting diode displays. Here, we investigate the physical mechanisms behind operational and environmental degradation of high-mobility, p-type polymer FETs and demonstrate an effective route to improve device stability. We show that water incorporated in nanometre-sized voids within the polymer microstructure is the key factor in charge trapping and device degradation. By inserting molecular additives that displace water from these voids, it is possible to increase the stability as well as uniformity to a high level sufficient for demanding industrial applications.