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Current laboratory diagnostic approaches for virus detection give reliable results, but they require a lengthy procedure, trained personnel, and expensive equipment and reagents; hence, they are not a suitable choice for home monitoring purposes. This paper addresses this challenge by developing a portable impedimetric biosensing system for the identification of COVID-19 patients. This sensing system has two main parts: a throwaway two-working electrode (2-WE) strip and a novel read-out circuit, specifically designed for simultaneous signal acquisition from both working electrodes. Highly reliable electrochemical signal tracking from multiplex immunosensors provides a potential for flexible and portable multi-biomarker detection. The electrodes' surfaces were functionalized with SARS-CoV-2 Nucleocapsid Antibody enabling the selective detection of Nucleocapsid protein (N-protein) along with self-validation in the clinical nasopharyngeal swab specimens. The proposed programmable highly sensitive impedance read-out system allows for a wide dynamic detection range, which makes the sensor capable of detecting N-protein concentrations between 0.116 and 10,000 pg/mL. This lightweight and economical read-out arrangement is an ideal prospect for being mass-produced, especially during urgent pandemic situations. Also, such an impedimetric sensing platform has the potential to be redesigned for targeting not only other infectious diseases but also other critical disorders.
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Manipulation of micro- and nanoparticles in complex biofluids is highly demanded in most biological and biomedical applications. A significant number of microfluidic platforms have been developed for inexpensive, rapid, accurate, and efficient particle manipulation. Due to the enormous potential of viscoelastic fluids (VEFs) for particle manipulation, various emerging microfluidic-based VEFs techniques have been presented over the last decade. This review provides an intuitive understanding of VEF physics for particle separation in different microchannel geometries. Besides, active and passive VEF methods are critically reviewed, highlighting the potential and practical challenges of each technique for particle/cell focusing, sorting, and separation. The outcome of this study could enable recognizing deliverable VEF technology with the promising prospect in the manipulation of submicron biological samples (e.g., exosomes, DNA, and proteins).
Assuntos
Técnicas Analíticas Microfluídicas , Nanopartículas/química , Nanotecnologia , Animais , Separação Celular/instrumentação , Células Cultivadas , Elasticidade , Desenho de Equipamento , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Nanotecnologia/instrumentação , Nanotecnologia/métodos , Tamanho da Partícula , ViscosidadeRESUMO
Organ-on-a-chip systems are miniaturized microfluidic 3D human tissue and organ models designed to recapitulate the important biological and physiological parameters of their in vivo counterparts. They have recently emerged as a viable platform for personalized medicine and drug screening. These in vitro models, featuring biomimetic compositions, architectures, and functions, are expected to replace the conventional planar, static cell cultures and bridge the gap between the currently used preclinical animal models and the human body. Multiple organoid models may be further connected together through the microfluidics in a similar manner in which they are arranged in vivo, providing the capability to analyze multiorgan interactions. Although a wide variety of human organ-on-a-chip models have been created, there are limited efforts on the integration of multisensor systems. However, in situ continual measuring is critical in precise assessment of the microenvironment parameters and the dynamic responses of the organs to pharmaceutical compounds over extended periods of time. In addition, automated and noninvasive capability is strongly desired for long-term monitoring. Here, we report a fully integrated modular physical, biochemical, and optical sensing platform through a fluidics-routing breadboard, which operates organ-on-a-chip units in a continual, dynamic, and automated manner. We believe that this platform technology has paved a potential avenue to promote the performance of current organ-on-a-chip models in drug screening by integrating a multitude of real-time sensors to achieve automated in situ monitoring of biophysical and biochemical parameters.
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Automação/métodos , Técnicas Biossensoriais/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Organoides/fisiologia , Automação/instrumentação , Técnicas Biossensoriais/instrumentação , Avaliação Pré-Clínica de Medicamentos/instrumentação , Coração/fisiologia , Humanos , Fígado/química , Fígado/fisiologia , Microfluídica , Modelos Biológicos , Miocárdio , Organoides/química , Organoides/efeitos dos fármacosRESUMO
Controlling stem cell (SC) fate is an extremely important topic in the realm of SC research. A variety of different external cues mainly mechanical, chemical, or electrical stimulations individually or in combination have been incorporated to control SC fate. Here, we will deconstruct the probable relationship between the functioning of electromagnetic (EMF) and SC fate of a variety of different SCs. The electromagnetic (EM) nature of the cells is discussed with the emphasis on the effects of EMF on the determinant factors that directly and/or indirectly influence cell fate. Based on the EM effects on a variety of cellular processes, it is believed that EMFs can be engineered to provide a controlled signal with the highest impact on the SC fate decision. Considering the novelty and broad applications of applying EMFs to change SC fate, it is necessary to shed light on many unclear mechanisms underlying this phenomenon.
Assuntos
Campos Eletromagnéticos , Células-Tronco/fisiologia , Animais , Ciclo Celular/fisiologia , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Campos Eletromagnéticos/efeitos adversos , Humanos , Mitocôndrias/metabolismo , Células-Tronco/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Tip-growing cells have the unique property of invading living tissues and abiotic growth matrices. To do so, they exert significant penetrative forces. In plant and fungal cells, these forces are generated by the hydrostatic turgor pressure. Using the TipChip, a microfluidic lab-on-a-chip device developed for tip-growing cells, we tested the ability to exert penetrative forces generated in pollen tubes, the fastest-growing plant cells. The tubes were guided to grow through microscopic gaps made of elastic polydimethylsiloxane material. Based on the deformation of the gaps, the force exerted by the elongating tubes to permit passage was determined using finite element methods. The data revealed that increasing mechanical impedance was met by the pollen tubes through modulation of the cell wall compliance and, thus, a change in the force acting on the obstacle. Tubes that successfully passed a narrow gap frequently burst, raising questions about the sperm discharge mechanism in the flowering plants.
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Dimetilpolisiloxanos/química , Técnicas Analíticas Microfluídicas/métodos , Tubo Polínico/química , Camellia/metabolismo , Elasticidade , Desenho de Equipamento , Análise de Elementos Finitos , Microfluídica/métodos , Modelos Biológicos , Plantas , Pressão , Estresse MecânicoRESUMO
The pollen tube is the most rapidly growing cell in the plant kingdom and has the function to deliver the sperm cells for fertilization. The growing tip region of the cell behaves in a chemotropic manner to respond to the guidance cues emitted by the pistil and the female gametophyte, but how it perceives and responds to these directional triggers is virtually unknown. Quantitative assessment of chemotropic behavior can greatly be enhanced by the administration of pharmacological or other biologically active agents at subcellular precision, which is a technical challenge when the target area moves as it grows. We developed a laminar flow based microfluidic device that allows for continuous administration of two different solutions with a movable interface that permits the dynamic targeting of the growing pollen tube apex over prolonged periods of time. Asymmetric administration of calcium revealed that rather than following the highest calcium concentration as would be expected with simple chemotropic behavior, the pollen tube of Camellia targets an optimal concentration suggesting the presence of two superimposed mechanisms. Subcellular application of pectin methyl esterase (PME), an enzyme that modifies the growth behavior by rigidifying the pollen tube cell wall, caused the tube to turn away from the agent - providing important evidence for a previously proposed conceptual model of the growth mechanism.
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Cálcio/metabolismo , Camellia/crescimento & desenvolvimento , Tubo Polínico/crescimento & desenvolvimento , Camellia/ultraestrutura , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Parede Celular/metabolismo , Fertilização , Flores/crescimento & desenvolvimento , Flores/ultraestrutura , Modelos Biológicos , Óvulo Vegetal/crescimento & desenvolvimento , Óvulo Vegetal/ultraestrutura , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tubo Polínico/ultraestrutura , PolinizaçãoRESUMO
Large-scale phenotyping of tip-growing cells such as pollen tubes has hitherto been limited to very crude parameters such as germination percentage and velocity of growth. To enable efficient and high-throughput execution of more sophisticated assays, an experimental platform, the TipChip, was developed based on microfluidic and microelectromechanical systems (MEMS) technology. The device allows positioning of pollen grains or fungal spores at the entrances of serially arranged microchannels equipped with microscopic experimental set-ups. The tip-growing cells (pollen tubes, filamentous yeast or fungal hyphae) may be exposed to chemical gradients, microstructural features, integrated biosensors or directional triggers within the modular microchannels. The device is compatible with Nomarski optics and fluorescence microscopy. Using this platform, we were able to answer several outstanding questions on pollen tube growth. We established that, unlike root hairs and fungal hyphae, pollen tubes do not have a directional memory. Furthermore, pollen tubes were found to be able to elongate in air, raising the question of how and where water is taken up by the cell. The platform opens new avenues for more efficient experimentation and large-scale phenotyping of tip-growing cells under precisely controlled, reproducible conditions.
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Sistemas Microeletromecânicos/instrumentação , Sistemas Microeletromecânicos/métodos , Tubo Polínico/citologia , Tubo Polínico/crescimento & desenvolvimento , Ar , Alumínio , Células Imobilizadas , Cobre , Desenho de Equipamento , Germinação , Ouro , Hifas/citologia , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Microscopia de Fluorescência , Fenótipo , Pólen/citologia , Resistência ao CisalhamentoRESUMO
A biocompatible polydimethylsiloxane (PDMS) biomicrofluidic platform is designed, fabricated and tested to study protuberance growth of single plant cells in a micro-vitro environment. The design consists of an inlet to introduce the cell suspension into the chip, three outlets to conduct the medium or cells out of the chip, a main distribution chamber and eight microchannels connected to the main chamber to guide the growth of tip growing plant cells. The test cells used here were pollen grains which produce cylindrical protrusions called pollen tubes. The goal was to adjust the design of the microfluidic network with the aim to enhance the uniformly distributed positioning of pollen grains at the entrances of the microchannels and to provide identical fluid flow conditions for growing pollen tubes along each microchannel. Computational fluid analysis and experimental testing were carried out to estimate the trapping efficiencies of the different designs.
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Microfluídica/instrumentação , Pólen/crescimento & desenvolvimento , Materiais Biocompatíveis/química , Camellia , Simulação por Computador , Dimetilpolisiloxanos , Desenho de Equipamento , Técnicas Analíticas Microfluídicas/métodos , Modelos Teóricos , Tubo Polínico/crescimento & desenvolvimentoRESUMO
Rapid and accurate quantification of metabolites in different bodily fluids is crucial for a precise health evaluation. However, conventional metabolite sensing methods, confined to centralized laboratory settings, suffer from time-consuming processes, complex procedures, and costly instrumentation. Introducing the MXene/nitrogen-doped electrochemically exfoliated graphene (MXene@N-EEG) nanocomposite as a novel biosensing platform in this work addresses the challenges associated with conventional methods, leveraging the concept of molecularly imprinted polymers (MIP) enables the highly sensitive, specific, and reliable detection of metabolites. To validate our biosensing technology, we utilize agmatine as a significant biologically active metabolite. The MIP biosensor incorporates electrodeposited Prussian blue nanoparticles as a redox probe, facilitating the direct electrical signaling of agmatine binding in the polymeric matrix. The MXene@N-EEG nanocomposite, with excellent metal conductivity and a large electroactive specific surface area, effectively stabilizes the electrodeposited Prussian blue nanoparticles. Furthermore, increasing the content of agmatine-imprinted cavities on the electrode enhances the sensitivity of the MIP biosensor. Evaluation of the designed MIP biosensor in buffer solution and plasma samples reveals a wide linear concentration range of 1.0 nM-100.0 µM (R2 = 0.9934) and a detection limit of 0.1 nM. Notably, the developed microfluidic biosensor offers low cost, rapid response time to the target molecule (10 min of sample incubation), good recovery results for detecting agmatine in plasma samples, and acceptable autonomous performance for on-chip detection. Moreover, its high reliability and sensitivity position this MIP-based biosensor as a promising candidate for miniaturized microfluidic devices with the potential for scalable production for point-of-care applications.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Grafite , Polímeros Molecularmente Impressos , Nanocompostos , Nitrogênio , Grafite/química , Técnicas Biossensoriais/métodos , Nanocompostos/química , Nitrogênio/química , Polímeros Molecularmente Impressos/química , Técnicas Eletroquímicas/métodos , Humanos , Limite de Detecção , Impressão Molecular , Polímeros/químicaRESUMO
Invasive behaviour is the hallmark of a variety of cell types of animal, plant, and fungal origin. Here we review the purpose and mechanism of invasive growth and migration. The focus is on the physical principles governing the process, the source of invasive force, and the cellular mechanism by which the cell penetrates the substrate. The current experimental methods for measuring invasive force and the modelling approaches for studying invasive behaviour are explained, and future experimental strategies are proposed.
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Parede Celular/fisiologia , Células Eucarióticas/fisiologia , Fungos/fisiologia , Fenômenos Fisiológicos Vegetais , Animais , Fenômenos Biomecânicos , Células Eucarióticas/citologia , Fungos/citologia , Fungos/crescimento & desenvolvimento , Mecanotransdução Celular , Modelos Biológicos , Desenvolvimento Vegetal , Raízes de Plantas/citologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/fisiologia , Tubo Polínico/citologia , Tubo Polínico/crescimento & desenvolvimento , Tubo Polínico/fisiologiaRESUMO
The separation of rare cells from complex biofluids has attracted attention in biological research and clinical applications, especially for cancer detection and treatment. In particular, various technologies and methods have been developed for the isolation of circulating tumor cells (CTCs) in the blood. Among them, the induced-charge electrokinetic (ICEK) flow method has shown its high efficacy for cell manipulation where micro-vortices (MVs), generated as a result of induced charges on a polarizable surface, can effectively manipulate particles and cells in complex fluids. While the majority of MVs have been induced by AC electric fields, these vortices have also been observed under a DC electric field generated around a polarizable hurdle. In the present numerical work, the capability of MVs for the manipulation of CTCs and their entrapment in the DC electric field is investigated. First, the numerical results are verified against the available data in the literature. Then, various hurdle geometries are employed to find the most effective geometry for MV-based particle entrapment. The effects of electric field strength (EFS), wall zeta potential magnitude, and the particles' diameter on the trapping efficacy are further investigated. The results demonstrated that the MVs generated around only the rectangular hurdle are capable of trapping particles as large as the size of CTCs. An EFS of about 75 V/cm was shown to be effective for the entrapment of above 90% of CTCs in the MVs. In addition, an EFS of 85 V/cm demonstrated a capability for isolating particles larger than 8 µm from a suspension of particles/cells 1-25 µm in diameter, useful for the enrichment of cancer cells and potentially for the real-time and non-invasive monitoring of drug effectiveness on circulating cancer cells in blood circulation.
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Stress affects cognition, behavior, and physiology, leading to lasting physical and mental illness. The ability to detect and measure stress, however, is poor. Increased circulating cortisol during stress is mirrored by cortisol release from sweat glands, providing an opportunity to use it as an external biomarker for monitoring internal emotional state. Despite the attempts at using wearable sensors for monitoring sweat cortisol, there is a lack of reliable wearable sweat collection devices that preserve the concentration and integrity of sweat biomolecules corresponding to stress levels. Here, a flexible, self-powered, evaporation-free, bubble-free, surfactant-free, and scalable capillary microfluidic device, MicroSweat, is fabricated to reliably collect human sweat from different body locations. Cortisol levels are detected corresponding to severe stress ranging from 25 to 125 ng mL-1 averaged across multiple body regions and 100-1000 ng mL-1 from the axilla. A positive nonlinear correlation exists between cortisol concentration and stress levels quantified using the perceived stress scale (PSS). Moreover, owing to the sweat variation in response to environmental effects and physiological differences, the longitudinal and personalized profile of sweat cortisol is acquired, for the first time, for various body locations. The obtained sweat cortisol data is crucial for analyzing human stress in personalized and clinical healthcare sectors.
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Suor , Dispositivos Eletrônicos Vestíveis , Humanos , Microfluídica , Hidrocortisona , Glândulas SudoríparasRESUMO
Elevated glial fibrillary acidic protein (GFAP) in the blood serum is one of the promising bodily fluid markers for the diagnosis of central nervous system (CNS) injuries, including traumatic brain injury (TBI), stroke, and spinal cord injury (SCI). However, accurate and point-of-care (POC) quantification of GFAP in clinical blood samples has been challenging and yet to be clinically validated against gold-standard assays and outcome practices. This work engineered and characterized a novel nanoporous carbon screen-printed electrode with significantly increased surface area and conductivity, as well as preserved stability and anti-fouling properties. This nano-decorated electrode was immobilized with the target GFAP antibody to create an ultrasensitive GFAP immunosensor and quantify GFAP levels in spiked samples and the serum of CNS injury patients. The immunosensor presented a dynamic detection range of 100 fg/mL to 10 ng/mL, a limit of detection of 86.6 fg/mL, and a sensitivity of 20.3 Ω mL/pg mm2 for detecting GFAP in the serum. Its clinical utility was demonstrated by the consistent and selective quantification of GFAP comparable to the ultrasensitive single-molecule array technology in 107 serum samples collected from TBI, stroke, and SCI patients. Comparing the diagnostic and prognostic performance of the immunosensor with the existing clinical paradigms confirms the immunosensor's accuracy as a potential complement to the existing imaging diagnostic modalities and presents a potential for rapid, accurate, cost-effective, and near real-time POC diagnosis and prognosis of CNS injuries.
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Técnicas Biossensoriais , Lesões Encefálicas Traumáticas , Nanoporos , Traumatismos da Medula Espinal , Acidente Vascular Cerebral , Humanos , Carbono , Proteína Glial Fibrilar Ácida , Biomarcadores , Imunoensaio , Lesões Encefálicas Traumáticas/diagnóstico , Traumatismos da Medula Espinal/diagnóstico , Acidente Vascular Cerebral/diagnósticoRESUMO
The evaporation of particle-laden sessile droplets is associated with capillary-driven outward flow and leaves nonuniform coffee-ring-like particle patterns due to far-from-equilibrium effects. Traditionally, the surface energies of the drop and solid phases are tuned, or external forces are applied to suppress the coffee-ring; however, achieving a uniform and repeatable particle deposition is extremely challenging. Here, we report a simple, scalable, and noninvasive technique that yields uniform and exceptionally ordered particle deposits on a microscale surface area by placing the droplet on a near neutral-wet shadow mold attached to a hydrophilic substrate. The simplicity of the method, no external forces, and no tuning materials' physiochemical properties make the present generic approach an excellent candidate for a wide range of sensitive applications. We demonstrate the utility of this method for fabricating ordered mono- and multilayer patternable coatings, producing nanofilters with controlled pore size, and creating reproducible functionalized nanosensors.
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The widespread and long-lasting effect of the COVID-19 pandemic has called attention to the significance of technological advances in the rapid diagnosis of SARS-CoV-2 virus. This study reports the use of a highly stable buffer-based zinc oxide/reduced graphene oxide (bbZnO/rGO) nanocomposite coated on carbon screen-printed electrodes for electrochemical immuno-biosensing of SARS-CoV-2 nuelocapsid (N-) protein antigens in spiked and clinical samples. The incorporation of a salt-based (ionic) matrix for uniform dispersion of the nanomixture eliminates multistep nanomaterial synthesis on the surface of the electrode and enables a stable single-step sensor nanocoating. The immuno-biosensor provides a limit of detection of 21 fg/mL over a linear range of 1-10â¯000 pg/mL and exhibits a sensitivity of 32.07 ohms·mL/pg·mm2 for detection of N-protein in spiked samples. The N-protein biosensor is successful in discriminating positive and negative clinical samples within 15 min, demonstrating its proof of concept used as a COVID-19 rapid antigen test.
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Antígenos Virais/análise , COVID-19/diagnóstico , Proteínas do Nucleocapsídeo de Coronavírus/análise , Grafite/química , Nanocompostos/química , Óxido de Zinco/química , Anticorpos Imobilizados/imunologia , Antígenos Virais/imunologia , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Humanos , Imunoensaio/instrumentação , Imunoensaio/métodos , Limite de Detecção , Fosfoproteínas/análise , Fosfoproteínas/imunologia , Estudo de Prova de Conceito , SARS-CoV-2/químicaRESUMO
The integration of electrochemical biosensors into fluid handling units such as paper-based, centrifugal, and capillary microfluidic devices has been explored with the purpose of developing point-of-care platforms for quantitative detection of bodily fluid markers. However, the present fluidic device designs largely lack the capacity of full assay automation, needing manual loading of one or multiple reagents or requiring external devices for liquid manipulation. Such fluidic handing platforms also require universality for detecting various biomarkers. These platforms are also largely produced using materials unsuitable for scalable manufacturing and with a high production cost. The mechanism of fluid flow also often induces noise to the embedded biosensors which adversely impacts the accuracy of biosensing. This work addresses these challenges by presenting a reliable design of a fully automated and universal capillary-driven microfluidic platform that automates several steps of label-free electrochemical biosensing assays. These steps include sample aliquoting, controlled incubation, removal of non-specific bindings, reagent mixing and delivery to sensing electrodes, and electrochemical detection. The multilayer architecture of the microfluidic device is made of polymeric and adhesive materials commercially used for the fabrication of point-of-care devices. The design and geometry of different components of the device (e.g., sampling unit, mixer, resistances, delay valves, interconnecting components) were optimized using a combined experimental testing and numerical fluid flow modeling to reach high reproducibility and minimize the noise-induced to the biosensor. As a proof of concept, the performance of this on-chip immunosensing platform was demonstrated for rapid and autonomous detection of glial fibrillary acidic proteins (GFAP) in phosphate-buffered saline (PBS). The microfluidic immunosensing device exhibited a linear detection range of 10-1000 pg mL-1 for the detection of GFAP within 30 min, with a limit of detection (LoD) and sensitivity of 3 pg mL-1 and 39 mL pg-1 mm-2 in PBS, respectively. Owing to its simplicity, sample-to-result performance, universality for handing different biofluids, low cost, high reproducibility, compatibility with scalable production, and short analysis time, the proposed biosensing platform can be further adapted for the detection of other biomarkers in different clinical bodily fluids for rapid diagnostic and prognostic applications.
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Técnicas Biossensoriais , Técnicas Analíticas Microfluídicas , Traumatismos do Sistema Nervoso , Biomarcadores , Proteína Glial Fibrilar Ácida , Humanos , Sistemas Automatizados de Assistência Junto ao Leito , Reprodutibilidade dos TestesRESUMO
There is a growing interest to understand if and how the gut microbiome is causally linked to the pathogenesis and/or progression of diseases. While in vitro cell line models are commonly used for studying specific aspects of the host-microbe interaction, gnotobiotic murine models are considered the preferred platform for studying causality in microbiome research. Nevertheless, findings from animal studies provide limited opportunity for delineating various areas of interest to the human gut microbiome research. Gut-on-chips are biomimetics recapitulating intestinal physiology which enable investigation of bidirectional effects of the host and microbiome. We posit that they could advance causal and ecological gut microbiome research in three major areas: (i) diet-microbiome and drug-microbiome interaction; (ii) microbiome-targeted therapeutics pharmacoecology; and (iii) mechanistic studies of gut microbiome and microbiome-targeted intervention in extraintestinal pathologies.
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Microbioma Gastrointestinal , Microbiota , Animais , Dieta , Microbioma Gastrointestinal/fisiologia , Interações entre Hospedeiro e Microrganismos , Humanos , CamundongosRESUMO
Electrochemical immuno-biosensors are one of the most promising approaches for accurate, rapid, and quantitative detection of protein biomarkers. The two-working electrode strip is employed for creating a self-supporting system, as a tool for self-validating the acquired results for added reliability. However, the realization of multiplex electrochemical point-of-care testing (ME-POCT) requires advancement in portable, rapid reading, easy-to-use, and low-cost multichannel potentiostat readers. The combined multiplex biosensor strips and multichannel readers allow for suppressing the possible complex matrix effect or ultra-sensitive detection of different protein biomarkers. Herein, a handheld binary-sensing (BiSense) bi-potentiostat was developed to perform electrochemical impedance spectroscopy (EIS)-based signal acquisition from a custom-designed dual-working-electrode immuno-biosensor. BiSense employs a commercially available microcontroller and out-of-shelf components, offering the cheapest yet accurate and reliable time-domain impedance analyzer. A specific electrical board design was developed and customized for impedance signal analysis of SARS-CoV-2 nucleocapsid (N)-protein biosensor in spiked samples and alpha variant clinical nasopharyngeal (NP) swab samples. BiSense showed limit-of-detection (LoD) down to 56 fg/mL for working electrode 1 (WE1) and 68 fg/mL for WE2 and reported with a dynamic detection range of 1 pg/mL to 10 ng/mL for detection of N-protein in spiked samples. The dual biosensing of N-protein in this work was used as a self-validation of the biosensor. The low-cost (â¼USD$40) BiSense bi-potentiostat combined with the immuno-biosensors successfully detected COVID-19 infected patients in less than 10 min, with the BiSense reading period shorter than 1.5 min, demonstrating its potential for the realization of ME-POCTs for rapid and hand-held diagnosis of infections.
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Técnicas Biossensoriais , COVID-19 , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Técnicas Eletroquímicas , Humanos , Reprodutibilidade dos Testes , SARS-CoV-2RESUMO
Multiplex electrochemical biosensors have been used for eliminating the matrix effect in complex bodily fluids or enabling the detection of two or more bioanalytes, overall resulting in more sensitive assays and accurate diagnostics. Many electrochemical biosensors lack reliable and low-cost multiplexing to meet the requirements of point-of-care detection due to either limited functional biosensors for multi-electrode detection or incompatible readout systems. We developed a new dual electrochemical biosensing unit accompanied by a customized potentiostat to address the unmet need for point-of-care multi-electrode electrochemical biosensing. The two-working electrode system was developed using screen-printing of a carboxyl-rich nanomaterial containing ink, with both working electrodes offering active sites for recognition of bioanalytes. The low-cost bi-potentiostat system (â¼$80) was developed and customized specifically to the bi-electrode design and used for rapid, repeatable, and accurate measurement of electrochemical impedance spectroscopy signals from the dual biosensor. This binary electrochemical data acquisition (Bi-ECDAQ) system accurately and selectively detected SARS-CoV-2 Nucleocapsid protein (N-protein) in both spiked samples and clinical nasopharyngeal swab samples of COVID-19 patients within 30 min. The two working electrodes offered the limit of detection of 116 fg/mL and 150 fg/mL, respectively, with the dynamic detection range of 1-10,000 pg/mL and the sensitivity range of 2744-2936 Ω mL/pg.mm2 for the detection of N-protein. The potentiostat performed comparable or better than commercial Autolab potentiostats while it is significantly lower cost. The open-source Bi-ECDAQ presents a customizable and flexible approach towards addressing the need for rapid and accurate point-of-care electrochemical biosensors for the rapid detection of various diseases.
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Técnicas Biossensoriais , COVID-19 , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Técnicas Eletroquímicas/métodos , Eletrodos , Humanos , Proteínas do Nucleocapsídeo , SARS-CoV-2RESUMO
Droplets produced within microfluidics have not only attracted the attention of researchers to develop complex biological, industrial and clinical testing systems but also played a role as a bit of data. The flow of droplets within a network of microfluidic channels by stimulation of their movements, trajectories, and interaction timing, can provide an opportunity for preparation of complex and logical microfluidic circuits. Such mechanical-based circuits open up avenues to mimic the logic of electrical circuits within microfluidics. Recently, simple microfluidic-based logical elements such as AND, OR, and NOT gates have been experimentally developed and tested to model basic logic conditions in laboratory settings. In this work, we develop new microfluidic networks, control the shape of channels and speed of droplet movement, and regulate the size of bubbles in order to extend the logical elements to six new logic gates, including AND/OR type 1, AND/OR type 2, NOT type 1, NOT type 2, Flip-Flop, Synchronizer, and a parametric model of T-junction as a bubble generator. We further designed and simulated a novel microfluidic Decoder 1 to 2, a Decoder 2 to 4, and a microfluidic circuit that combines several individual logic gates into one complex circuit. Further fabrication and experimental testing of these newly introduced logic gates within microfluidics enable implementing complex circuits in high-throughput microfluidic platforms for tissue engineering, drug testing and development, and chemical synthesis and process design.