Biomedical Microdevices最新文献

Respiratory viruses can cause epidemics or pandemics, which are worldwide outbreaks of disease. The severity of these events varies depending on the virus, its characteristics, along with environmental factors. The frequency of epidemics and pandemics caused by respiratory viruses is difficult to predict, but the potential severity of such events underlines the importance of continued monitoring, research, and preparation for emerging infectious diseases. To help improve pandemic preparedness, we created a fully integrated duplex reverse transcription loop-mediated isothermal amplification (RT-LAMP) device targeting two respiratory viruses, influenza A/X-31 virus and bovine coronavirus, as a replacement for SARS-CoV-2. This device can be adapted to any other respiratory virus. In this study, we showed and evaluated a prototype of a microfluidic system, and showed that duplex RT-LAMP can detect and distinguish between the two viruses, with LoDs of 2,000 copies/ml for bovine coronavirus and 200 copies/ml for influenza A/X-31 virus.

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Microelectrode arrays (MEAs) have proven to be a powerful tool to study electrophysiological processes over the last decades with most technology developed for investigation of the heart or brain. Other targets in the field of bioelectronic medicine are the peripheral nervous system and its innervation of various organs. Beyond the heart and nervous systems, the beta cells of the pancreatic islets of Langerhans generate action potentials during the production of insulin. In vitro experiments have demonstrated that their activity is a biomarker for blood glucose levels, suggesting that recording their activity in vivo could support patients suffering from diabetes mellitus with long-term automated read-out of blood glucose concentrations. Here, we present a flexible polymer-based implant having 64 low impedance microelectrodes designed to be implanted to a depth of 10 mm into the pancreas. As a first step, the implant will be used in acute experiments in pigs to explore the electrophysiological processes of the pancreas in vivo. Beyond use in the pancreas, our flexible implant and simple implantation method may also be used in other organs such as the brain.

Accurate, rapid, and multiplexed nucleic acid detection is critical for environmental and biomedical monitoring. In recent years, CRISPR-Cas12a has shown great potential in improving the performance of DNA biosensing. However, the nonspecific trans-cleavage activity of Cas12a complicates the multiplexing capability of Cas12a biosensing. We report a 3D-printed composable microfluidic plate (cPlate) device that utilizes miniaturized wells and microfluidic loading for a multiplexed CRISPR-Cas12a assay. The device easily combines loop-mediated isothermal amplification (LAMP) and CRISPR-Cas12a readout in a simple and high-throughput workflow with low reagent consumption. To ensure the maximum performance of the device, the concentration of Cas12a and detection probe was optimized, which yielded a four-fold sensitivity improvement. Our device demonstrates sensitive detection to the fg mL^− 1 level for four waterborne pathogens including shigella, campylobacter, cholera, and legionella within 1 h, making it suitable for low-resource settings.

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Cytobrushes are used for low-invasive sample collection and screening in multiple diseases, with a significant impact on early detection, prevention, and diagnosis. This study focuses on improving the safety of cell brushing in hard-to-reach locations by exploring brush construction from absorbable materials. We investigated the efficacy of loop brushes made of absorbable suture wires of Chirlac, Chirasorb, Monocryl, PDS II, Vicryl Rapid, Glycolon, and Catgut during their operation in conjunction with fine-needle aspiration in an artificial cyst model. PDS II brushes demonstrated the highest efficiency, while Monocryl and Catgut also provided a significant brushing effect. Efficient brushes portrayed higher flexural rigidity than their counterparts, and their efficiency was inversely proportional to their plastic deformation by the needle. Our results open avenues for safer cell biopsies in hard-to-reach locations by utilizing brushes composed of absorbable materials.

Colorimetric assays rely on detecting colour changes to measure the concentration of target molecules. Paper substrates are commonly used for the detection of biomarkers due to their availability, porous structure, and capillarity. However, the morphological and mechanical properties of paper, such as fibre diameter, pore size, and tensile strength, cannot be easily tuned to meet the specific requirements of colorimetric sensors, including liquid capacity and reagent immobilisation. As an alternative to paper materials, biodegradable polymeric membranes made of electrospun polycaprolactone (PCL) fibres can provide various tunable properties related to fibre diameter and pore size.

We aimed to obtain a glucose sensor substrate for colorimetric sensing using electrospinning with PCL. A feeding solution was created by mixing PCL/chloroform and 3,3’,5’,5’-tetramethylbenzidine (TMB)/ethanol solutions. This solution was electrospun to fabricate a porous membrane composed of microfibres consist of PCL and TMB. The central area of the membrane was made hydrophilic through air plasma treatment, and it was subsequently functionalized with a solution containing glucose oxidase, horseradish peroxidase, and trehalose.

The sensing areas were evaluated by measuring colour changes in glucose solutions of varying concentrations. The oxidation reactions of glucose and TMB in sensor substrates were recorded and analysed to establish the correlation between different glucose concentrations and colour changes. For comparison, conventional paper substrates prepared with same parameters were evaluated alongside the electrospun PCL substrates. As a result, better immobilization of reagents and higher sensitivity of glucose were achieved with PCL substrates, indicating their potential usage as a new sensing substrate for bioassays.

Local field potentials, the extracellular electrical activities from brain regions, provide clinically relevant information about the status of neurophysiological conditions, including epilepsy. In this study, a 13-channel silicon-based single-shank microelectrode array (MEA) was designed and fabricated to record local field potentials (LFPs) from the different depths of a rat’s brain. A titanium/gold layer was patterned as electrodes on an oxidized silicon substrate, and silicon dioxide was deposited as a passivation layer. The fabricated array was implanted in the somatosensory cortex of the right hemisphere of an anesthetized rat. The developed MEA was interfaced with an OpenBCI Cyton Daisy Biosensing Board to acquire the local field potentials. The LFPs were acquired at three different neurophysiological conditions, including baseline signals, chemically-induced epileptiform discharges, and recovered baseline signals after anti-epileptic drug (AED) administration. Further, time-frequency analyses were performed on the acquired biopotentials to study the difference in spatiotemporal features. The processed signals and time-frequency analyses clearly distinguish between pre-convulsant and post-AED baselines and evoked epileptiform discharges.

Whole mount zebrafish antibody staining (ABS) is a common staining technique used to localize protein information in a zebrafish embryo or larva. Like most biological assays, the whole mount zebrafish ABS is still largely conducted manually through labor intensive and time-consuming steps which affect both consistency and throughput of the assay. In this work, we develop a milli fluidic device that can automatically trap and immobilize the fixed chorion-less zebrafish embryos for the whole mount ABS. With just a single loading step, the zebrafish embryos can be trapped by the milli fluidic device through a chaotic hydrodynamic trapping process. Moreover, a consistent body orientation (i.e., head point inward) for the trapped zebrafish embryos can be achieved without additional orientation adjustment device. Furthermore, we employed a consumer-grade SLA 3D printer assisted method for device prototyping which is ideal for labs with limited budgets. Notably, the milli fluidic device has enabled the optimization and successful implementation of whole mount zebrafish Caspase-3 ABS. We demonstrated our device can accelerate the overall procedure by reducing at least 50% of washing time in the standard well-plate-based manual procedure. Also, the consistency is improved, and manual steps are reduced using the milli fluidic device. This work fills the gap in the milli fluidic application for whole mount zebrafish immunohistochemistry. We hope the device can be accepted by the zebrafish community and be used for other types of whole mount zebrafish ABS procedures or expanded to more complicated in situ hybridization (ISH) procedure.

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The association of machine learning (ML) tools with the synthesis of nanoparticles has the potential to streamline the development of more efficient and effective nanomedicines. The continuous-flow synthesis of nanoparticles via microfluidics represents an ideal playground for ML tools, where multiple engineering parameters – flow rates and mixing configurations, type and concentrations of the reagents – contribute in a non-trivial fashion to determine the resultant morphological and pharmacological attributes of nanomedicines. Here we present the application of ML models towards the microfluidic-based synthesis of liposomes loaded with a model hydrophobic therapeutic agent, curcumin. After generating over 200 different liposome configurations by systematically modulating flow rates, lipid concentrations, organic:water mixing volume ratios, support-vector machine models and feed-forward artificial neural networks were trained to predict, respectively, the liposome dispersity/stability and size. This work presents an initial step towards the application and cultivation of ML models to instruct the microfluidic formulation of nanoparticles.

Creating micrometer-resolution high-aspect-ratio three-dimensional (3D) structures remain very challenging despite significant microfabrication methods developed for microelectromechanical systems (MEMS). This is especially the case when such structures are desired to be metallic to support electronic applications. Here, we present a microfabrication process that combines two-photon-polymerization (2PP) printing to create a polymeric high-aspect-ratio three-dimensional structure and electroless metal plating that selectively electroplates only the polymeric structure to create high-aspect-ratio 3D metallic structures having micrometer-resolution. To enable this, the effect of various 2PP processing parameters on SU-8 photoresist microstructures were first systematically studied. These parameters include laser power, slicing/hatching distances, and pre-/post-baking temperature. This optimization resulted in a maximum aspect ratio (height to width) of ~ 12. Following this polymeric structure printing, electroless plating using Tollens’ Reagent were utilized to selectively coat silver particles only on the polymeric structure, but not on the silicon substrate. The final 3D metallic structures were evaluated in terms of their resistivity, reproducibly showing resistivity of ~ 10^–6 [Ω·m]. The developed 3D metallic structure microfabrication process can be further integrated with conventional 2D lithography to achieve even more complex structures. The developed method overcomes the limitations of current MEMS fabrication processes, allowing a variety of previously impossible metallic microstructures to be created.

Early diagnosis of C reactive protein (CRP) is critical to applying effective therapies for related diseases. Diagnostic technology in today's healthcare systems is mostly deployed in central laboratories, involves expensive and time-consuming processes, and is operated by specialized personnel. For example, the enzyme-linked immunosorbent assay (ELISA), considered the gold standard diagnostic method, is labor-intensive and requires complex procedures such as multiple washing and labeling steps. Due to these limitations of current diagnostic techniques, it is difficult for people to regularly monitor their health and ultimately the disease is more likely to be diagnosed at a later stage. The problem is exacerbated for economically disadvantaged people living in underdeveloped countries. To address these challenges in the traditional diagnostic field, point-of-care (POC) biosensors have emerged as a promising alternative. This allows patients to have their health checked regularly at or near their bedside without resorting to laboratory tests. Nanotechnology-based methods such as biosensors have been extensively researched and developed. Among biosensors, there are also label-free biosensors with high sensitivity that do not require complicated procedures and reduce test time. However, some drawbacks such as high cost, bulky size and need for trained personnel to operate have not been improved. In this review article, we provide an overview of routine methods in CRP diagnosis and then introduce biosensors as a modern, advanced alternative to older methods. Readers of this article can learn about biosensing and its benefits while being aware of the limitations of routine methods.

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