Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H2O2 depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes.
Traumatic brain injury (TBI) has been identified as an important cause of death and severe disability in all age groups and particularly in children and young adults. Central to TBI’s devastation is a delayed secondary injury that occurs in 30-40% of TBI patients each year, while they are in the hospital Intensive Care Unit (ICU). Secondary injuries reduce survival rate after TBI and usually occur within 7 days post-injury. State-of-art monitoring of secondary brain injuries benefits from the acquisition of high-quality and time-aligned electrical data i.e. ElectroCorticoGraphy (ECoG) recorded by means of strip electrodes placed on the brain’s surface, and neurochemical data obtained via rapid sampling microdialysis and microfluidics-based biosensors measuring brain tissue levels of glucose, lactate and potassium. This article progresses the field of multi-modal monitoring of the injured human brain by presenting the design and realisation of a new, compact, medical-grade amperometry, potentiometry and ECoG recording bioinstrumentation. Our combined TBI instrument enables the high-precision, real-time neuroelectrochemical monitoring of TBI patients, who have undergone craniotomy neurosurgery and are treated sedated in the ICU. Electrical and neurochemical test measurements are presented, confirming the high-performance of the reported TBI bioinstrumentation.
Capillary pumps are integral components of passive microfluidic devices. They can displace precise volumes of liquid, avoiding the need for external active components, providing a solution for sample preparation modules in Point-of-Care (PoC) diagnostic platforms. In this work, we describe a variety of high-performance capillary pump designs, suitable for the Lab-on-Printed-Circuit-Board technology (LoPCB). Pumps are fabricated entirely on Printed Circuit Board (PCB) substrates via commercially available manufacturing processes. We demonstrate the concept of LoPCB technology and detail the fabrication method of different architectures of PCB-based capillary pumps. The capillary pumps are combined with microfluidic channels of various hydraulic resistances and characterised experimentally for different micropillar shapes and minimum feature size. Their performance in terms of flow rate is reported. Due to the superhydrophilic properties of oxygen plasma treated FR-4 PCB substrate, the capillary pump flow rates are much higher (138 μL/min, for devices comprising micropillar arrays without preceding microchannel) than comparable devices based on glass, silicon or polymers. Finally, we comment on the technology’s prospects, such as incorporating more complicated microfluidic networks that can be tailored for assays.
27 October 2016 in Annals of Biomedical Engineering
Exposure of endothelial cells to low and multidirectional blood flow is known to promote a pro-atherogenic phenotype. The mechanics of the vessel wall is another important mechano-stimulus within the endothelial cell environment, but no study has examined whether changes in the magnitude and direction of cell stretch can be pro-atherogenic. Herein, we developed a custom cell stretching device to replicate the in vivo stretch environment of the endothelial cell and examined whether low and multidirectional stretch promote nuclear translocation of NF-κB. A fluid–structure interaction model of the device demonstrated a nearly uniform strain within the region of cell attachment and a negligible magnitude of shear stress due to cyclical stretching of the cells in media. Compared to normal cyclical stretch, a low magnitude of cyclical stretch or no stretch caused increased expression of nuclear NF-κB (p = 0.09 and p < 0.001, respectively). Multidirectional stretch also promoted significant nuclear NF-κB expression, comparable to the no stretch condition, which was statistically higher than the low (p < 0.001) and normal (p < 0.001) stretch conditions. This is the first study to show that stretch conditions analogous to atherogenic blood flow profiles can similarly promote a pro-atherogenic endothelial cell phenotype, which supports a role for disturbed vessel wall mechanics as a pathological cell stimulus in the development of advanced atherosclerotic plaques.
A novel, Lab-on-Printed Circuit Board (LoPCB) manufacturing technology is demonstrated for the development of low-cost electrochemical biosensors combined with microfluidics for Point-of-Care (PoC) applications. An analysis of the developed PCB architecture is presented, detailing the three development areas of the proposed LoPCB platform, i.e. microfluidics, biosensors and electronics. Design rules and potential fabrication limitations are also discussed, based on the characterization of prototype fabricated systems. Two PCB-based devices have been designed and fabricated, a microfluidic active diluter with a variable and actively controlled dilution ratio and an electrochemical biosensor. The obtained results demonstrate the feasibility of a complete LoPCB platform, where all three compartments will co-exist and co-operate, providing an electronic-based PoC system for electrochemical biosensing.
We introduce a compact time-domain system for near-infrared spectroscopy using a spread spectrum technique. The proof-of-concept single channel instrument utilises a low-cost commercially available optical transceiver module as a light source, controlled by a Kintex 7 field programmable gate array (FPGA). The FPGA modulates the optical transceiver with maximum-length sequences at line rates up to 10Gb/s, allowing us to achieve an instrument response function with full width at half maximum under 600ps. The instrument is characterised through a set of detailed phantom measurements as well as proof-of-concept in vivo measurements, demonstrating performance comparable with conventional pulsed time-domain near-infrared spectroscopy systems.