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The limit of detection (LOD), speed, and cost of crucial COVID-19 diagnostic tools, including lateral flow assays (LFA), enzyme-linked immunosorbent assays (ELISA), and polymerase chain reactions (PCR), have all improved because of the financial and governmental support for the epidemic. The most notable improvement in overall efficiency among them has been seen with PCR. Its significance for human health increased during the COVID-19 pandemic, when it emerged as the commonly used approach for identifying the virus. However, because of problems with speed, complexity, and expense, PCR deployment in point-of-care settings continues to be difficult. Microfluidic platforms offer a promising solution by enabling the development of smaller, more affordable, and faster PCR systems. In this review, we delve into the engineering challenges associated with the advancement of high-speed microfluidic PCR equipment. We introduce criteria that facilitate the evaluation and comparison of factors such as speed, LOD, cycling efficiency, and multiplexing capacity, considering sample volume, fluidics, PCR reactor geometry and materials, as well as heating/cooling methods. We also provide a comprehensive list of commercially available PCR devices and conclude with projections and a discussion regarding the current obstacles that need to be addressed in order to progress further in this field.
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Técnicas Biossensoriais , COVID-19 , Humanos , COVID-19/diagnóstico , Pandemias , Reação em Cadeia da Polimerase , Microfluídica , Teste para COVID-19RESUMO
Membranes are fundamental elements within organ-on-a-chip (OOC) platforms, as they provide adherent cells with support, allow nutrients (and other relevant molecules) to permeate/exchange through membrane pores, and enable the delivery of mechanical or chemical stimuli. Through OOC platforms, physiological processes can be studied in vitro, whereas OOC membranes broaden knowledge of how mechanical and chemical cues affect cells and organs. OOCs with membranes are in vitro microfluidic models that are used to replace animal testing for various applications, such as drug discovery and disease modeling. In this review, the relevance of OOCs with membranes is discussed as well as their scaffold and actuation roles, properties (physical and material), and fabrication methods in different organ models. The purpose was to aid readers with membrane selection for the development of OOCs with specific applications in the fields of mechanistic, pathological, and drug testing studies. Mechanical stimulation from liquid flow and cyclic strain, as well as their effects on the cell's increased physiological relevance (IPR), are described in the first section. The review also contains methods to fabricate synthetic and ECM (extracellular matrix) protein membranes, their characteristics (e.g., thickness and porosity, which can be adjusted depending on the application, as shown in the graphical abstract), and the biological materials used for their coatings. The discussion section joins and describes the roles of membranes for different research purposes and their advantages and challenges.
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Fabrication of highly aligned fibers by far-field electrospinning is a challenging task to accomplish. Multiple studies present advances in the alignment of electrospun fibers which involve modification of the conventional electrospinning setup with complex additions, multi-phased fabrication, and expensive components. This study presents a new collector design with an origami structure to produce highly-aligned far-field electrospun fibers. The origami collector mounts on the rotating drum and can be easily attached and removed for each round of fiber fabrication. This simple, effective, and inexpensive technique yields high-quality ultra-aligned fibers while the setup remains intact for other fabrication types. The electrospun poly(É-caprolactone) (PCL) fibers were assessed by scanning electron microscope (SEM), fiber diameter distribution, water contact angle (WCA), Fast Fourier Transform analysis (FFT), surface plot profile, and pixel intensity plots. We thoroughly explored the impact of influential parameters, including polymer concentration, injection rate, collector rotation speed, distance from the collector to the tip, and needle gauge number on fibers' quality and alignment. Moreover, we employed machine learning algorithms to predict the outcomes and classify the high-quality fibers instead of low-quality productions.
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Many advanced microfluidic Lab-on-disc (LOD) devices require an on-board power supply for powering active components. LODs with an on-board electrical power supply are called electrified-LODs (eLODs) and are the subject of the present review. This survey comprises two main parts. First, we discuss the different means of delivering electrical energy to a spinning disc including slip-ring, wireless power transmission, and on-board power supply. In the second part, we focus on utilizing electrical power on eLODs for three electrokinetic microfluidic processes: electrophoresis, electroosmotic flow, and dielectrophoresis. Electrokinetic phenomena enable propulsion, separation, and manipulation of different fluids and various types of microparticles/cells. We summarize the theoretical and experimental results for all three electrokinetic phenomena enacted on centrifugal platforms. While extensive numerical modeling and experimental research are available for electrokinetics on stationary platforms, there is a noticeable lack of development in this area when executed on rotating platforms. The review concludes by comparing the strengths and weaknesses of different electrokinetic techniques implemented on centrifugal platforms, and additionally, the most promising applications of electrokinetic-assisted eLOD devices are singled out.
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Técnicas Biossensoriais , Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas/métodos , Eletro-Osmose , Eletroforese , Técnicas Analíticas Microfluídicas/normas , Técnicas Analíticas Microfluídicas/tendênciasRESUMO
In recent years, wearable contact lenses for medical applications have attracted significant attention, as they enable continuous real-time recording of physiological information via active and noninvasive measurements. These devices play a vital role in continuous monitoring of intraocular pressure (IOP), noninvasive glucose monitoring in diabetes patients, drug delivery for the treatment of ocular illnesses, and colorblindness treatment. In specific, this class of medical devices is rapidly advancing in the area of drug loading and ocular drug release through incorporation of electrospun fibers. The electrospun fiber matrices offer a high surface area, controlled morphology, wettability, biocompatibility, and tunable porosity, which are highly desirable for controlled drug release. This article provides an overview of the advances of contact lens devices in medical applications with a focus on four main applications of these soft wearable devices: (i) IOP measurement and monitoring, (ii) glucose detection, (iii) ocular drug delivery, and (iv) colorblindness treatment. For each category and application, significant challenges and shortcomings of the current devices are thoroughly discussed, and new areas of opportunity are suggested. We also emphasize the role of electrospun fibers, their fabrication methods along with their characteristics, and the integration of diverse fiber types within the structure of the wearable contact lenses for efficient drug loading, in addition to controlled and sustained drug release. This review article also presents relevant statistics on the evolution of medical contact lenses over the last two decades, their strengths, and the future avenues for making the essential transition from clinical trials to real-world applications.
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The survival of living beings, including humanity, depends on a continuous supply of clean water. However, due to the development of industry, agriculture, and population growth, an increasing number of wastewaters is discarded, and the negative effects of such actions are clear. The first step in solving this situation is the collection and monitoring of pollutants in water bodies to subsequently facilitate their treatment. Nonetheless, traditional sensing techniques are typically laboratory-based, leading to potential diminishment in analysis quality. In this paper, the most recent developments in micro- and nano-electrochemical devices for pollutant detection in wastewater are reviewed. The devices reviewed are based on a variety of electrodes and the sensing of three different categories of pollutants: nutrients and phenolic compounds, heavy metals, and organic matter. From these electrodes, Cu, Co, and Bi showed promise as versatile materials to detect a grand variety of contaminants. Also, the most commonly used material is glassy carbon, present in the detection of all reviewed analytes.
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Poluentes Ambientais , Metais Pesados , Nanoestruturas , Técnicas Eletroquímicas , Eletrodos , Poluentes Ambientais/análise , Metais Pesados/análise , Nanoestruturas/química , ÁguaRESUMO
In this study, we carried out a heterogeneous cytoplasmic lipid content screening of Neochloris oleoabundans microalgae by dielectrophoresis (DEP), using castellated glassy carbon microelectrodes in a PDMS microchannel. For this purpose, microalgae were cultured in nitrogen-replete (N+) and nitrogen-deplete (N-) suspensions to promote low and high cytoplasmic lipid production in cells, respectively. Experiments were carried out over a wide frequency window (100 kHz-30 MHz) at a fixed amplitude of 7 VPP. The results showed a statistically significant difference between the dielectrophoretic behavior of N+ and N- cells at low frequencies (100-800 kHz), whereas a weak response was observed for mid- and high frequencies (1-30 MHz). Additionally, a finite element analysis using a 3D model was conducted to determine the dielectrophoretic trapping zones across the electrode gaps. These results suggest that low-cost glassy carbon is a reliable material for microalgae classification-between low and high cytoplasmic lipid content-through DEP, providing a fast and straightforward mechanism.
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Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a particular coronavirus strain responsible for the coronavirus disease 2019 (COVID-19), accounting for more than 3.1 million deaths worldwide. Several health-related strategies have been successfully developed to contain the rapidly-spreading virus across the globe, toward reduction of both disease burden and infection rates. Particularly, attention has been focused on either the development of novel drugs and vaccines, or by adapting already-existing drugs for COVID-19 treatment, mobilizing huge efforts to block disease progression and to overcome the shortage of effective measures available at this point.Areas covered: This perspective covers the breakthrough of multifunctional biomimetic cell membrane-based nanoparticles as next-generation nanosystems for cutting-edge COVID-19 therapeutics and vaccination, specifically cell membrane-derived nanovesicles and cell membrane-coated nanoparticles, both tailorable cell membrane-based nanosystems enriched with the surface repertoire of native cell membranes, toward maximized biointerfacing, immune evasion, cell targeting and cell-mimicking properties.Expert opinion: Nano-based approaches have received widespread interest regarding enhanced antigen delivery, prolonged blood circulation half-life and controlled release of drugs. Cell membrane-based nanoparticles comprise interesting antiviral multifunctional nanoplatforms for blocking SARS-CoV-2 binding to host cells, reducing inflammation through cytokine neutralization and improving drug delivery toward COVID-19 treatment.
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Tratamento Farmacológico da COVID-19 , Nanopartículas , Antivirais/uso terapêutico , Membrana Celular , Humanos , SARS-CoV-2 , VacinaçãoRESUMO
We demonstrate a loop-mediated isothermal amplification (LAMP) method to detect and amplify SARS-CoV-2 genetic sequences using a set of in-house designed initiators that target regions encoding the N protein. We were able to detect and amplify SARS-CoV-2 nucleic acids in the range of 62 to 2 × 105 DNA copies by this straightforward method. Using synthetic SARS-CoV-2 samples and RNA extracts from patients, we demonstrate that colorimetric LAMP is a quantitative method comparable in diagnostic performance to RT-qPCR (i.e., sensitivity of 92.85% and specificity of 81.25% in a set of 44 RNA extracts from patients analyzed in a hospital setting).
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Teste de Ácido Nucleico para COVID-19/métodos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA/análise , SARS-CoV-2/química , Carga Viral/métodos , COVID-19/diagnóstico , Colorimetria/métodos , Proteínas do Nucleocapsídeo de Coronavírus , DNA/análise , DNA/química , Corantes Fluorescentes/química , Humanos , Substâncias Intercalantes/química , Fenolsulfonaftaleína/química , Fosfoproteínas , RNA/químicaRESUMO
Cancer is one of the leading causes of annual deaths worldwide, accounting for nearly 10 million deaths each year. Metastasis, the process by which cancer spreads across the patient's body, is the main cause of death in cancer patients. Because the rising trend observed in statistics of new cancer cases and cancer-related deaths does not allow for an optimistic viewpoint on the future-in relation to this terrible disease-the scientific community has sought methods to enable early detection of cancer and prevent the apparition of metastatic tumors. One such method is known as liquid biopsy, wherein a sample is taken from a bodily fluid and analyzed for the presence of CTCs or other cancer biomarkers (e.g., growth factors). With this objective, interest is growing by year in electrokinetically-driven microfluidics applied for the concentration, capture, filtration, transportation, and characterization of CTCs. Electrokinetic techniques-electrophoresis, dielectrophoresis, electrorotation, and electrothermal and EOF-have great potential for miniaturization and integration with electronic instrumentation for the development of point-of-care devices, which can become a tool for early cancer diagnostics and for the design of personalized therapeutics. In this contribution, we review the state of the art of electrokinetically-driven microfluidics for cancer cells manipulation.
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Biomarcadores Tumorais , Eletroforese , Técnicas Analíticas Microfluídicas , Células Neoplásicas Circulantes , Células Tumorais Cultivadas , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Humanos , Dispositivos Lab-On-A-Chip , Neoplasias/diagnóstico , Neoplasias/patologia , Neoplasias/terapia , Células Neoplásicas Circulantes/química , Células Neoplásicas Circulantes/metabolismo , Células Tumorais Cultivadas/química , Células Tumorais Cultivadas/citologia , Células Tumorais Cultivadas/metabolismoRESUMO
Pyrolysis is a feasible solution for environmental problems related to the inadequate disposal of waste tires, as it leads to the recovery of pyrolytic products such as carbon black, liquid fuels and gases. The characteristics of pyrolytic carbon black can be enhanced through chemical activation in order to produce the required properties for its application. In the search to make the waste tire pyrolysis process profitable, new applications of the pyrolytic solid products have been explored, such as for the fabrication of energy-storage devices and precursor in the synthesis of nanomaterials. In this study, waste tires powder was chemically activated using acid (H2SO4) and/or alkali (KOH) to recover pyrolytic carbon black with different characteristics. H2SO4 removed surface impurities more thoroughly, improving the carbon black's surface area, while KOH increased its oxygen content, which improved the carbon black's stability in water suspension. Pyrolytic carbon black was fully characterized by elemental analysis, inductively coupled plasma-optical emission spectrometry (ICP-OES), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, X-ray diffraction (XRD), N2 adsorption/desorption, scanning electron microscopy-energy-dispersive X-ray spectroscopy (SEM-EDS), dynamic light scattering (DLS), and ζ potential measurement. In addition, the pyrolytic carbon black was used to explore its feasibility as a precursor for the synthesis of carbon dots; synthesized carbon dots were analyzed preliminarily by SEM and with a fluorescence microplate reader, revealing differences in their morphology and fluorescence intensity. The results presented in this study demonstrate the effect of the activating agent on pyrolytic carbon black from waste tires and provide evidence of the feasibility of using waste tires for the synthesis of nanomaterials such as carbon dots.
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The classic theory of direct-current (DC) insulator-based dielectrophoresis (iDEP) considers that, in order to elicit particle trapping, dielectrophoretic (DEP) velocity counterbalances electrokinetic (EK) motion, that is, electrophoresis (EP) and electro-osmotic flow (EOF). However, the particle velocity DEP component requires empirical correction factors (sometimes as high as 600) to account for experimental observations, suggesting the need for a refined model. Here, we show that, when applied to particle suspensions, a high-magnitude DC uniform electric field induces nonlinear particle velocities, leading to particle flow reversal beyond a critical field magnitude, referred to as the EK equilibrium condition. We further demonstrate that this particle motion can be described through an exploratory induced-charge EP nonlinear model. The model predictions were validated under an insulator-based microfluidic platform demonstrating predictive particle trapping for three different particle sizes (with an estimation error < 10%, not using correction factors). Our findings suggest that particle motion and trapping in "DC-iDEP" devices are dominated by EP and EOF, rather than by DEP effects.
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The current global health threat by the novel coronavirus disease 2019 (COVID-19) requires an urgent deployment of advanced therapeutic options available. The role of nanotechnology is highly relevant to counter this "virus" nano enemy. Nano intervention is discussed in terms of designing effective nanocarriers to counter the conventional limitations of antiviral and biological therapeutics. This strategy directs the safe and effective delivery of available therapeutic options using engineered nanocarriers, blocking the initial interactions of viral spike glycoprotein with host cell surface receptors, and disruption of virion construction. Controlling and eliminating the spread and reoccurrence of this pandemic demands a safe and effective vaccine strategy. Nanocarriers have potential to design risk-free and effective immunization strategies for severe acute respiratory syndrome coronavirus 2 vaccine candidates such as protein constructs and nucleic acids. We discuss recent as well as ongoing nanotechnology-based therapeutic and prophylactic strategies to fight against this pandemic, outlining the key areas for nanoscientists to step in.
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Infecções por Coronavirus/prevenção & controle , Vacinação em Massa/métodos , Nanotecnologia/métodos , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Vacinas Virais/uso terapêutico , COVID-19 , Vacinas contra COVID-19 , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/terapia , Infecções por Coronavirus/virologia , Humanos , Vacinação em Massa/efeitos adversos , Pneumonia Viral/imunologia , Pneumonia Viral/terapia , Pneumonia Viral/virologia , Vacinas Virais/imunologiaRESUMO
The use of multiphase flows in microfluidics to carry dispersed phase material (droplets, particles, bubbles, or fibers) has many applications. In this review paper, we focus on such flows on centrifugal microfluidic platforms and present different methods of dispersed phase material generation. These methods are classified into three specific categories, i.e., step emulsification, crossflow, and dispenser nozzle. Previous works on these topics are discussed and related parameters and specifications, including the size, material, production rate, and rotational speed are explicitly mentioned. In addition, the associated theories and important dimensionless numbers are presented. Finally, we discuss the commercialization of these devices and show a comparison to unveil the pros and cons of the different methods so that researchers can select the centrifugal droplet/particle generation method which better suits their needs.
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Polymer solutions with different concentrations of SU-8 2002/poly(ethylene) glycol/tetrabutyl ammonium tetrafluoroborate (SU-8/PEO/TBATFB) were electrospun in a low-voltage near-field electrospinning platform (LVNFES) at different velocities. Their diameters were related to the concentration contents as well as to their Deborah (De) numbers, which describes the elasticity of the polymer solution under determined operating conditions. We found a direct correlation between the concentration of PEO/TBATFB, the De and the diameter of the fibers. Fibers with diameters as thin as 465 nm can be achieved for De ≈ 1.
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This paper introduces the concept of continuous chaotic printing, i.e. the use of chaotic flows for deterministic and continuous extrusion of fibers with internal multilayered micro- or nanostructures. Two free-flowing materials are coextruded through a printhead containing a miniaturized Kenics static mixer (KSM) composed of multiple helicoidal elements. This produces a fiber with a well-defined internal multilayer microarchitecture at high-throughput (>1.0 m min-1). The number of mixing elements and the printhead diameter determine the number and thickness of the internal lamellae, which are generated according to successive bifurcations that yield a vast amount of inter-material surface area (â¼102 cm2 cm-3) at high resolution (â¼10 µm). This creates structures with extremely high surface area to volume ratio (SAV). Comparison of experimental and computational results demonstrates that continuous chaotic 3D printing is a robust process with predictable output. In an exciting new development, we demonstrate a method for scaling down these microstructures by 3 orders of magnitude, to the nanoscale level (â¼150 nm), by feeding the output of a continuous chaotic 3D printhead into an electrospinner. The simplicity and high resolution of continuous chaotic printing strongly supports its potential use in novel applications, including-but not limited to-bioprinting of multi-scale layered biological structures such as bacterial communities, living tissues composed of organized multiple mammalian cell types, and fabrication of smart multi-material and multilayered constructs for biomedical applications.
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Bioimpressão , Nanoestruturas/química , Alginatos/química , Bactérias/citologia , Grafite/química , Reprodutibilidade dos Testes , Engenharia TecidualRESUMO
Centrifugal microfluidic platforms or lab-on-discs (LODs) have evolved into a popular technology for automating chemical and biological assays. LODs today enable scientists to implement and integrate different operational units, including fluid mixing, droplet generation, cell-sorting, gene amplification, analyte detection, and so forth. For an efficient design and cost-effective implementation of any microfluidic device, including LODs, theoretical analysis and considerations should play a more important role than they currently do. The theoretical analysis we will show is especially essential to the investigation of detailed phenomena at the small length scales and high-speed typical for LODs where a wide range of forces may be involved. Previous LOD review papers presented mostly experimental results with theory as an afterthought. Hence, a review paper focused on the theoretical aspects, and associated computational studies of LOD devices is an urgent need. In the present review paper, all previous computational studies on LOD devices are categorized as single-phase flows, two-phase flows, network simulation, and solids. For each of these categories, the governing equations and important formulas are presented and explained. Moreover, a handy scaling analysis is introduced to aid scientists when comparing different competing forces in LOD devices. We hope that by surveying and contrasting various theoretical LOD studies, we shed some light on existing controversies and reveal where additional theoretical work is needed.
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Based on the concept of LEGO toys, a fiber probe analytical platform (FPAP) was developed as a powerful diagnostic tool offering higher sensitivity in detection of infectious agents compared to established methods. Using the form and the function of LEGO toys, this protocol describes a fiber-based, 96-well plate, which suspends a new class of chemically-designed, electrospun fibers within the assay. This clamping strategy allows both sides of the developed fiber mats to interact with biomolecules within the assay thus benefiting from the tailored chemical and physical properties of these fiber-based bioreceptors in attracting the biomolecules to the surface. The fabrication method of FPAP involves one-step electrospinning of the chemically designed fibers, 3D printing of the LEGO-like probing segments, and assembly of the device followed by ELISA procedure. FPAP follows the same principles of operation as that of a conventional enzyme linked immunosorbent assay (ELISA), therefore, it can be run by lab technicians, expert in ELISA. FPAP was used for early diagnosis of Dengue fever and provided an 8-fold higher sensitivity while the limit of detection (LOD) was recorded to be in femto-gram per milliliter range which is significantly low when compared to other existing techniques or conventional assay. This platform allows different types of paper/fiber bio-receptive platforms to be incorporated within the design that promises simultaneous recognition of multiple infectious agents.
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Anticorpos Antivirais , Vírus da Dengue/imunologia , Dengue , Imunoglobulina G , Impressão Tridimensional , Anticorpos Antivirais/química , Anticorpos Antivirais/imunologia , Dengue/diagnóstico , Dengue/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/química , Imunoglobulina G/imunologia , Limite de DetecçãoRESUMO
We herein report a high-resolution nanopatterning method using low voltage electromechanical spinning with a rotating collector to obtain aligned graphitized micro and nanowires for carbon nanomanufacturing. A small wire diameter and a small inter-wire spacing were obtained by controlling the electric field, the spinneret-to-collector distance, the pyrolysis parameters, the linear speed of the spinneret, the rotational speed of the collector. Using a simple scaling analysis, we show how the straightness and the diameter of the wires can be controlled by the electric field and the distance of the spinneret to the collector. A small inter-wire spacing, as predicted by a simple model, was achieved by simultaneously controlling the linear speed of the spinneret and the rotational speed of the collector. Rapid drying of the polymer nanowires enabled the facile fabrication of suspended wires over various structures. Patterned polyacrylonitrile wires were carbonized using standard stabilization and pyrolysis to obtain carbon nanowires. Suspended carbon nanowires with a diameter of <50 nm were obtained. We also established a method for making patterned, highly graphitized structures by using the aforementioned carbon wire structures as a template for chemical vapor deposition of graphite. This patterning technique offers high throughput for nano writing, which outperforms other existing nanopatterning techniques, making it a potential candidate for large-scale carbon nanomanufacturing.
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Glassy carbon nanofibers (GCNFs) are considered promising candidates for the fabrication of nanosensors for biosensing applications. Importantly, in part due to their great stability, carbon electrodes with sub-10 nm nanogaps represent an attractive platform for probing the electrical characteristics of molecules. The fabrication of sub-10 nm nanogap electrodes in these GCNFs, which is achieved by electrically stimulating the fibers until they break, was previously found to require fibers shorter than 2 µm; however, this process is generally hampered by the limitations inherent to photolithographic methods. In this work, to obtain nanogaps on the order of 10 nm without the need for sub-2 µm GCNFs, we employed a fabrication strategy in which the fibers were gradually thinned down by continuously monitoring the changes in the electrical resistance of the fiber and adjusting the applied voltage accordingly. To further reduce the nanogap size, we studied the mechanism behind the thinning and eventual breakdown of the suspended GCNFs by controlling the environmental conditions and pressure during the experiment. Following this approach, which includes performing the experiments in a high-vacuum chamber after a series of carbon dioxide (CO2) purging cycles, nanogaps on the order of 10 nm were produced in suspended GCNFs 52 µm in length, much longer than the ~2 µm GCNFs needed to produce such small gaps without the procedure employed in this work. Furthermore, the electrodes showed no apparent change in their shape or nanogap width after being stored at room temperature for approximately 6 months.