Microfluidic Paper-based Devices Research Articles

Printed Capillary Microfluidic Devices and Their Application in Biosensing.

Microfluidic devices with a free-standing structure were printed directly on polymer films using the functional materials that form interconnected pores. The printed devices can transport fluids by capillary action in the same fashion as paper-based microfluidic devices, and they can handle much smaller sample volumes than typical paper-based devices. Detection of glucose was performed using both colorimetric and electrochemical methods, and the observed limits of detection (LOD) were similar to those obtained with paper-based microfluidic devices under comparable testing conditions. It is demonstrated that printed microfluidic devices can be fabricated using printing processes that are suitable for high-volume and low-cost production and that the integration of microfluidic channels with electrodes is straightforward with printing. Several materials that are printable and form interconnected pores are presented.

Salivary calcium determination with a specially developed microfluidic paper-based device for point-of-care analysis

The calcium monitoring in the body not only anticipates several potential diseases (osteoporosis, kidney stones or high blood pressure) but also helps to improve target therapies and follow-up the patient's health status. Calcium monitoring is essential for the diagnosis of one of the most common endocrine disorders worldwide, namely hyperparathyroidism. So, in this work, a new Point-of-care test (POC-test) using a microfluidic paper-based analytical device (μPAD) for calcium quantification in saliva samples is described. The developed μPAD was based on the colorimetric reaction between calcium and cresolphthalein complexone (CPC) which forms an intense purple colour product. The developed device enabled calcium quantification in the range of 0.27–4.50 mmol/L (11.0–180 mg/L) with a detection limit of 80 µmol/L (3.2 mg/L). The accuracy of the developed μPAD was confirmed by analysing saliva samples (#10) and comparing the results obtained with the atomic absorption spectrometry reference procedure; the relative deviation between the two sets of results was below 10 %. A correlation between salivary calcium content and calcium content in blood was established and it was possible to conclude that salivary calcium concentrations above 1.55 mmol/L is an indicator of hypercalcemia. The developed device was stable for 2 weeks when stored at room temperature in vacuum conditions.

Electroanalytical paper-based device for reliable detection and quantification of sugars in milk

Knowing the level of sugars in food and beverages is essential because they are related to diseases such as diabetes, and obesity, among others. In this regard, an approach has been designed, characterized, and applied to develop the first all-in-one paper-based microfluidic analytical device for the quantification of sugars in milk samples. In particular, the powerful combination of the paper-based platform, the facile manufacturing, and the on-demand customization, allowed to obtain a stand-alone platform that was capable to store all the reagents necessary for the assay, to treating the sample by removing major interferences due to lipids and proteins, and to visualize the results in minutes. A wax-patterned chromatographic paper was finely designed to provide a reagent-free platform for the use of non-specialists applied towards the analysis of sugars' content (lactose as the main sugar). Sugars were electrochemically quantified down to 0.16 g L−1 with a repeatability of 5%. To strengthen the efficacy of the novel platform, the electrochemical paper-based analytical device has been applied to commercial milk, including skimmed, semi-skimmed, whole, and lactose-free, confirming satisfactory performance with analytical reliability. This paper-based architecture opens novel avenues for portable (bio)sensing of other relevant species, not only in the agri-food field.

µPED based electrode modified with nanocomposite for the detection of proline: An abiotic stress biomarker in plant

Plants when exposed to stressful conditions, accumulates array of metabolites. Among them proline plays significant and beneficial role as osmolyte, metal chelator and as anti-oxidative defense and regulator molecule. Thus the proline content is one of the potent bio-marker for the early detection of abiotic stress in plant. In this work a microfluidic paper-based electrochemical device (µPED) modified with copper oxide nanoparticles (CuO) decorated reduced graphene oxide (rGO) nanocomposite is utilized to develop disposable system for the detection of proline. Wax and screen printings were utilized to create microfluidic chambers and electrode pattern on paper device, respectively. Portable potentiostat was used for measuring the current in response from the addition of proline. The proposed sensor exhibits excellent electrochemical performance with broad linear responses over a proline concentration ranging from 0.1 mM to 7.5 mM (R2 =0.99) and showed lower detection limit of 0.05 mM at 0.4 V vs. pseudo Ag/AgCl. Excellent electrochemical performance may be attributed to presence of electroactive rGO and catalytic activity towards proline is due to the coordination between proline and copper (II) oxide nanoparticles. These results demonstrate that the proposed paper-based electrochemical sensor can be utilized in disposable and portable way to detect proline content in the abiotic-stressed plants and has potential to be applied in on-site application.

Paper-Based All-in-One Origami Nanobiosensor for Point-of-Care Detection of Cardiac Protein Markers in Whole Blood.

Rapid and accurate diagnosis of cardiovascular diseases (CVDs) at the earliest stage is of paramount importance to improve the treatment outcomes and avoid irreversible damage to a patient's cardiovascular system. Microfluidic paper-based devices (μPADs) represent a promising platform for rapid CVD diagnosis at the point of care (POC). This paper presents an electrochemical μPAD (E-μPAD) with an all-in-one origami design for rapid and POC testing of cardiac protein markers in whole blood. Based on the label-free, electrochemical impedance spectroscopy (EIS) immunoassay, the E-μPAD integrates all essential components on a single chip, including three electrochemical cells, a plasma separation membrane, and a buffer absorption pad, enabling easy and streamlined operations for multiplexed detection of three cardiac protein markers [cardiac troponin I (cTnI), brain natriuretic peptide (BNP)-32, and D-Dimer] on a finger-prick whole blood sample within 46 min. Superior analytical performance is achieved through sensitive EIS measurement on carbon electrodes decorated with semiconductor zinc oxide nanowires (ZnO NWs). Using spiked human plasma samples, ultralow limits of detection (LODs) of E-μPAD are achieved at 4.6 pg/mL (190 fM) for cTnI, 1.2 pg/mL (40 fM) for BNP-32, and 146 pg/mL (730 fM) for D-Dimer. Real human blood samples spiked with purified proteins are also tested, and the device's analytical performance was proven to be comparable to commercial ELISA kits. The all-in-one E-μPAD will allow rapid and sensitive testing of cardiac protein markers through easy operations, which holds great potential for on-site screening of acute CVDs in nonlaboratory settings such as emergency rooms, doctor's offices, or patient homes.

Paper-based microfluidic device for selective detection of peanut allergen Ara h1 applying black phosphorus-Au nanocomposites for signal amplification

This paper developed a portable microfluidic paper-based analysis device (μ-PAD) combined with the electrochemical technique for efficient and sensitive detection of peanut allergen Ara h1. The proposed μ-PAD works based on the variation of differential pulse voltammetry (DPV) response current induced by peanut allergen Ara h1. Black phosphorus (BP)–Au nanocomposites were introduced both to improve the electron transfer rate at the electrode interface for signal amplification, and to immobilize the specific Ara h1 aptamers through Au–S bonding to recognize the target in food matrices. This μ-PAD had good specificity and detection stability for Ara h1 allergen and could complete the entire analysis process within 20 min, achieving a wide linear response range (25–800 ng mL−1) and a low detection limit (LOD, 11.8 ng mL−1). In the Ara h1 allergen detection applied to real peanut products (cookies, milk, and bread), the constructed μ-PAD obtained acceptable recoveries (93.50%–101.86%) with relative standard deviations (RSDs) of 0.36–2.97% (n = 3), with a good correlation with the ELISA results (R2 = 0.9956). Therefore, the portable μ-PAD based on BP-Au nanocomposites was demonstrated to provide an effective strategy for rapid analysis and screening of Ara h1 allergen in food, which has broad application prospects.

Point-of-Care Devices for Viral Detection: COVID-19 Pandemic and Beyond.

The pandemic of COVID-19 and its widespread transmission have made us realize the importance of early, quick diagnostic tests for facilitating effective cure and management. The primary obstacles encountered were accurately distinguishing COVID-19 from other illnesses including the flu, common cold, etc. While the polymerase chain reaction technique is a robust technique for the determination of SARS-CoV-2 in patients of COVID-19, there arises a high demand for affordable, quick, user-friendly, and precise point-of-care (POC) diagnostic in therapeutic settings. The necessity for available tests with rapid outcomes spurred the advancement of POC tests that are characterized by speed, automation, and high precision and accuracy. Paper-based POC devices have gained increasing interest in recent years because of rapid, low-cost detection without requiring external instruments. At present, microfluidic paper-based analysis devices have garnered public attention and accelerated the development of such POCT for efficient multistep assays. In the current review, our focus will be on the fabrication of detection modules for SARS-CoV-2. Here, we have included a discussion on various strategies for the detection of viral moieties. The compilation of these strategies would offer comprehensive insight into the detection of the causative agent preparedness for future pandemics. We also provide a descriptive outline for paper-based diagnostic platforms, involving the determination mechanisms, as well as a commercial kit for COVID-19 as well as their outlook.

Focusing the intervention of paper-based microfluidic devices for the forensic investigative purposes

Focusing the intervention of paper-based microfluidic devices for the forensic investigative purposes

A portable multi-channel fluorescent paper-based microfluidic chip based on smartphone imaging for simultaneous detection of four heavy metals

Due to the excessive contamination of heavy metals pollution, it is very urgent and necessary to develop a real-time detection method for the heavy metals in food. As a target sensing device, a paper-based microfluidic device (μPAD) has the advantages of simplicity, low-cost, and portability. In this study, a self-driven microfluidic paper-based chip was first developed for the simultaneous detection of four targets. The channels on the microfluidic chip were prepared by using wax printing and automatic screen printing on the filter paper, where liquid flowed by capillary force without pump assistance. Based on the specific binding ability of aptamers to heavy metals, a “turn-on” fluorescence aptasensor for the simultaneous detection of four heavy metal ions was developed on the proposed multi-channel device via smartphone imaging. The obtained fluorescence images were digitized into RGB color values by Image J software, and an M-mode was established to realize the quantitative detection of heavy metal ions. Under optimal conditions, the limits of detection for lead(II), mercury(II), cadmium(II), and arsenic(III) were 4.20 nM, 1.70 nM, 2.04 nM, and 1.65 nM, respectively. Furthermore, the aptasensor was successfully applied to the quantitative detection of four heavy metal ions in apple and lettuce samples with recovery rates of 84.0%–104.1%.

Hierarchical Capillarity-Assisted Liquid Invasion in Multilayered Paper Channels for Nanoelectrokinetic Preconcentration.

A nanoelectrokinetic phenomenon called ion concentration polarization (ICP) has been recently applied to microfluidic paper-based devices for the high fold preconcentration of low-abundant analytes. The inherent microstructural characteristics of cellulose papers can sufficiently stabilize the chaotic electroconvection of ICP, which is a significant annoyance for typical engineered microfluidic channels. However, a high electrical voltage to induce ICP in a paper-fluidic channel can increase unavoidable electrophoretic forces over drag forces so that the preconcentrated plug is rapidly receded with severe dispersion. In order to enhance the hydraulic drag force that helps the preconcentration of analytes, here we introduce a multilayered paper structure into paper-fluidic channel. We theoretically and experimentally demonstrate that a hierarchical capillary structure in a multilayered paper-fluidic channel can effectively increase the hydraulic drag force. For the practical utility in the field of diagnostics, the mechanism is verified by a simple example of the immunoassay using biotin-streptavidin complexation.

Development of a biodegradable microfluidic paper-based device for blood-plasma separation integrated with non-enzymatic electrochemical detection of ascorbic acid

In the present article, we developed an electrochemical microfluidic paper-based device (EμPAD) for the non-enzymatic detection of Ascorbic Acid (AA) concentration in plasma using whole human blood. We combined LF1 blood plasma separation membrane and Whatman grade 1 filter paper to separate plasma from whole blood through wax printing. A screen-printed electrode (SPE) was modified with spherical-shaped MgFe2O4 nanomaterial (n-MgF) to improve the catalytic properties of SPE. The n-MgF was prepared via hydrothermal method, and its material phase and morphology were confirmed via XRD, FTIR, TEM, SEM, and AFM analysis. The fabricated n-MgF/SPE/EμPAD exhibited detection of AA ranging from 0 to 80 μM. The obtained value of the detection limit, limit of quantification, sensitivity, and response time are 2.44 μM, 8.135 μM, 5.71 × 10−3 mA μM−1 cm−2, and 10 s, respectively. Our developed n-MgF/SPE/EμPAD shows marginal interference with the common analytes present in plasma, such as uric acid, glutamic acid, glucose, urea, lactic acid, and their mixtures. Overall, our low-cost, portable device with its user-friendly design and efficient plasma separation capability offers a practical and effective solution for estimating AA concentration from whole human blood in a single step.

Functional toner for office laser printer and its application for printing of paper-based superwettable patterns and devices

Laserjet printing is a kind of facile and digital do-it-yourself strategies, which is of importance to fabricate inexpensive paper-based microfluidic devices. However, the printed hydrophobic barrier is not hydrophobic enough due to the weak hydrophobicity and requires subsequent heating, which can lead to the pyrolysis of cellulose in the paper and influence the detection results. Here, for the first time, we report a kind of functional toner including toner and polydopamine (PDA) nanocapsules which contains oleic acid modified ferric tetroxide (OA-Fe3O4) and octadecylamine (ODA), which is suitable for printing with desired shapes and sizes to lead to formation of superhydrophobic barriers. Moreover, patterns printed with functional toner have good stability, including resistance to moisture, ultraviolet (UV) and bending. Finally, a proof-of-concept of metal and nitrite ions testing is demonstrated using colorimetric analysis, and the results show that the printed devices successfully perform instant detection of ions. The developed functional toner offers easy fabrication, cost-effectiveness and mass production of paper-based devices. In general, this strategy provides a new idea and technical support for the rapid prototyping of microfluidic paper-based analytical devices (μPADs) using laserjet printing.

Application of Paper-Based Microfluidic Analytical Devices (µPAD) in Forensic and Clinical Toxicology: A Review.

The need for providing rapid and, possibly, on-the-spot analytical results in the case of intoxication has prompted researchers to develop rapid, sensitive, and cost-effective methods and analytical devices suitable for use in nonspecialized laboratories and at the point of need (PON). In recent years, the technology of paper-based microfluidic analytical devices (μPADs) has undergone rapid development and now provides a feasible, low-cost alternative to traditional rapid tests for detecting harmful compounds. In fact, µPADs have been developed to detect toxic molecules (arsenic, cyanide, ethanol, and nitrite), drugs, and drugs of abuse (benzodiazepines, cathinones, cocaine, fentanyl, ketamine, MDMA, morphine, synthetic cannabinoids, tetrahydrocannabinol, and xylazine), and also psychoactive substances used for drug-facilitated crimes (flunitrazepam, gamma-hydroxybutyric acid (GHB), ketamine, metamizole, midazolam, and scopolamine). The present report critically evaluates the recent developments in paper-based devices, particularly in detection methods, and how these new analytical tools have been tested in forensic and clinical toxicology, also including future perspectives on their application, such as multisensing paper-based devices, microfluidic paper-based separation, and wearable paper-based sensors.

On-hand tool for ammonium and urea determination in saliva to monitor chronic kidney disease – Design of a couple of microfluidic paper-based devices

In this work, two microfluidic paper-based analytical devices (μPADs) were developed for the quantification of urea and NHx in human saliva to aid in the diagnosis/monitoring of chronic kidney disease (CKD). The NHx determination was based on the conversion of ammonium to ammonia, followed by its diffusion through a hydrophobic membrane and then the color change of bromothymol blue (BTB) indicator. In the urea determination, prior to the ammonium conversion and BTB color change, the enzymatic conversion of urea into ammonium was produced, using urease. Several optimization studies were carried out to attain a quantification range of 0.10–5.0 mM with 0.032 mM limit of detection for the NHx μPAD, and a determination range of 0.16–5.0 mM with 0.049 mM limit of detection for the urea μPAD. The method accuracy was assessed, and the measurements obtained with NHx μPAD were compared with the ones obtained from an ammonia ion selective electrode; while the measurements of the urea μPAD were compared with the ones obtained from a commercially available kit. There were no statistically significant differences between methods, proving that both NHx and urea μPAD were effective on-hand tools for CKD monitoring in saliva. To evaluate their functionality as point-of-care devices, stability studies were also performed and revealed that both NHx and urea μPAD were stable when stored in a vacuum for 2 and 1 month, respectively. After the sample introduction, the NHx μPAD could be scanned within the first 2 h and the urea μPAD within 1 h.

Microfluidic paper-based device coupled with 3D printed imaging box for colorimetric detection in resource-limited settings

Rapid and effective methods for the detection of analytes such as water contaminants, food adulterants and biomolecules are essential for the protection of public health and environmental protection. Most of the currently established analytical techniques need sophisticated equipment, centralized testing facilities, costly operations, and trained personnel. Such limitations make them inaccessible to the general populace, particularly in regions with limited resources. The emergence of microfluidic devices offers a promising alternative to overcome several such constraints. This work describes a protocol for fabricating a low-cost, open-source paper-based microfluidic device using easily available tools and materials for colorimetric detection of analytes. The ease and simplicity of fabrication allow users to design customized devices. The device is coupled with an imaging box assembled from 3D printed parts to maintain uniform lighting conditions during analytical testing. The platform allows digital imaging using smartphones or cameras to instantaneously capture images of reaction zones on the device for quantitative analysis. The system is demonstrated for detecting hexavalent chromium, a toxic water contaminant. The image analysis is performed using open-source ImageJ for quantification of results. The approach demonstrated in this work can be readily adopted for a wide range of sensing applications.

Paper-based biosensors as point-of-care diagnostic devices for the detection of cancers: a review of innovative techniques and clinical applications.

The development and rapid progression of cancer are major social problems. Medical diagnostic techniques and smooth clinical care of cancer are new necessities that must be supported by innovative diagnostic methods and technologies. Current molecular diagnostic tools based on the detection of blood protein markers are the most common tools for cancer diagnosis. Biosensors have already proven to be a cost-effective and accessible diagnostic tool that can be used where conventional laboratory methods are not readily available. Paper-based biosensors offer a new look at the world of analytical techniques by overcoming limitations through the creation of a simple device with significant advantages such as adaptability, biocompatibility, biodegradability, ease of use, large surface-to-volume ratio, and cost-effectiveness. In this review, we covered the characteristics of exosomes and their role in tumor growth and clinical diagnosis, followed by a discussion of various paper-based biosensors for exosome detection, such as dipsticks, lateral flow assays (LFA), and microfluidic paper-based devices (µPADs). We also discussed the various clinical studies on paper-based biosensors for exosome detection.

Disposable Microfluidic Paper-Based Device for On-Site Quantification of Urinary Creatinine

In this work, a new microfluidic paper-based analytical device (µPAD) was developed for on-hand creatinine quantification in urine samples. When compared to conventional methods, this innovative paper device is more accessible and portable, it provides low-cost analysis (cost of consumables of 40 cents), and it is applicable to non-invasive biological fluids. Furthermore, the paper-based approach is used within an environmentally friendly assembly with no need for wax printing and small amounts of reagents resulting in low waste production and easy disposal by incineration. Its assembly method includes cutting paper discs arranged into several reading units within a plastic pouch, enabling effective creatinine quantification with accuracy based on a vertical flow approach. The method is based on the colourimetric reaction between creatinine and alkaline picric acid, where the solution colour changes from yellow to orange/red. Under optimal conditions, the developed method allowed creatinine quantification in the dynamic range of 2.20–35.0 mg/dL, with a limit of detection (LOD) of 0.66 mg/dL and a limit of quantification (LOQ) of 2.20 mg/dL. The colour intensity developed was processed in ImageJ software, based on digital image scanning, performed in 20 min (up to 4 h) after the sample insertion. The device is stable for up to one week when stored in a vacuum at 4 °C. The method was validated by comparing the results with a batch-wise procedure, where there were no statistically significant differences between both methods.

A 3D sliding-strip microfluidic device for the simultaneous determination of multiple target analytes

A simple paper-based microfluidic device was fabricated to simultaneously detect multiple targets. Microfluidic paper-based analytical devices (μPAD) comprise a single-layer moving sliding PAD (SPAD) to control the flow channel switch together with a folding origami PAD (OPAD) to test the target analytes. The facile assembly without any splicing materials avoids cross-contamination and non-specific adsorption of joining materials that may be caused by multi-target detection. The concentration of Fe(III), Ni(II), Cr(VI), and nitrite in standard solutions and actual aqueous solutions was successfully determined using the designed μPAD. The μPAD was able to achieve LOD of 3.3 mg/L, 1.3 mg/L, 0.35 mg/L, 0.28 mg/L for Fe (III), Ni (II), Cr (VI), and nitrite, respectively. The designed SOPAD exhibits improved stability, with a deviation of less than 7% compared to conventional analytical methods (ICP-OES and UV). Our work demonstrates that this 3D PAD holds great promise and a wide scope in environmental monitoring, biochemical analysis, food testing and other testing industries.

Paper-based microfluidics and tailored gold nanoparticles for visual colorimetric detection of multiplex allergens

The highly efficient and accurate recognition of targeted allergens is an essential element in the diagnosis of allergic diseases and follow-up desensitization treatment in clinic. The current clinical methods widely used to detect sIgE are high cost, time-consuming procedures, and bulky equipment. Herein, a multiplex microfluidic paper-based device (multi-μPAD) was developed that combined with tailored gold nanoparticles for simultaneously visual, colorimetric detection of different allergens in serum. This device could be used as quantitative detection of sIgE with LOD as low as 0.246 KUA/L in colorimetric method. In vitro results also showed that this device possessed good repeatability, high accuracy and incredible stability in different pH (6.0–7.4) and temperature (24–37 °C), as well as long-term storage within 90-day. Finally, this method was successfully utilized for assessing clinical multi-sample screening in 35 allergic patients. After the addition of the samples from allergic patients, the agreement rate of clinical results with commercial enzyme-linked immunosorbent assay (ELISA) kit reached more than 97%, which further indicated that this device had the advantages of efficient, accurate and sensitive to screen various allergens in real clinical serum samples. Therefore, by simply altering antigens and antibodies, this device can also be used for high-throughput detection of other allergens, making it considerable potential for clinical diagnosis of allergic diseases.

Solid state fluid transport and sensing based on printed and embedded cellulose-polymer composites: An alternative pathway to paper-based microfluidic devices

Solid state fluid transport and sensing based on printed and embedded cellulose-polymer composites: An alternative pathway to paper-based microfluidic devices