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Microfluidic paper-based analytical devices (μPADs) and microfluidic thread-based analytical devices (μTADs) have recently been introduced as a new class of on-site monitoring devices. Creating hydrophilic channels with hydrophobic barriers on papers/threads produces μPADs/μTADs. Fabrication is a crucial step in creating durable μPADs/μTADs that can withstand various liquids and impact the device's performance. Fabrication materials with distinct physicochemical properties allow microfluidic systems with sophisticated functions to be customized for specific applications. We present flexible and low-cost fabrication methods for μPAD and μTAD platforms. Platform designs and fabrications were implemented using a trial-and-error method for various designs with varying parameters. All production methods presented in the method section were used in |PAD production. For comparison studies, only the dipping method was used in μTAD production due to its ease of application. In this study, we tried to reveal the strengths and weaknesses of the production techniques and the resulting microfluidic platforms. A leaching test was performed with water solutions containing red ink. The compatibility of the hydrophobic walls of the platforms was tested with several solvents (isopropanol, methanol, and acetone), deionized (DI) water, and phosphate buffer solution PBS and compared. Patterning paper with polydimethylsiloxane (PDMS), white glue, alkyl ketene dimer (AKD), beeswax, and paraffin are much more flexible and simpler than traditional photoresist-based fabrications. The advantages and disadvantages of fabrication techniques; solvent resistance and wicking behaviors of platforms were discussed in the last part. The fabricated microfluidic platforms can be functionalized and used in many areas where analytical tests are applied. Studies on diversifying channel geometries and increasing resolution need to be continued. It should be investigated which devices can be used to obtain qualitative and quantitative results. To make simple and cheap production techniques suitable for mass production, studies should be carried out from different branches. [ABSTRACT FROM AUTHOR]

Microfluidic Paper‐based Analytical Devices (µPADs) have emerged as a new class of microfluidic systems, offering numerous advantages over traditional microfluidic chips. These advantages include simplicity, cost‐effectiveness, stability, storability, disposability, and portability. As a result, various designs for different types of assays are developed and investigated. In recent years, µPADs are combined with conventional detection methods to enable rapid on‐site detection, providing results comparable to expensive and sophisticated large‐scale testing methods that require more time and skilled personnel. The application of µPAD techniques is extensive in environmental quality control/analysis, clinical diagnosis, and food safety testing, paving the way for on‐site real‐time diagnosis as a promising future development. This review focuses on the recent research advancements in the design, fabrication, material selection, and detection methods of µPADs. It provides a comprehensive understanding of their principles of operation, applications, and future development prospects. [ABSTRACT FROM AUTHOR]

Iodine is an essential micronutrient for humans due to its fundamental role in the biosynthesis of thyroid hormones. As a key parameter to assess health conditions, iodine intake needs to be monitored to ascertain and prevent iodine deficiency. Iodine is available from various food sources (such as seaweed, fish, and seafood, among others) and dietary supplements (multivitamins or mineral supplements). In this work, a microfluidic paper-based analytical device (μPAD) to quantify iodide in seaweed and dietary supplements is described. The developed μPAD is a small microfluidic device that emerges as quite relevant in terms of its analytical capacity. The quantification of iodide is based on the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) by hydrogen peroxide in the presence of iodine, which acts as the catalyst to produce the blue form of TMB. Additionally, powder silica was used to intensify and uniformize the colour of the obtained product. Following optimization, the developed μPAD enabled iodide quantification within the range of 10–100 µM, with a detection limit of 3 µM, and was successfully applied to seaweeds and dietary supplements. The device represents a valuable tool for point-of-care analysis, can be used by untrained personnel at home, and is easily disposable, low-cost, and user-friendly. [ABSTRACT FROM AUTHOR]

Cannabis is recorded to be the most used drug in the world. This causes a lot of difficulties for developing countries to curb the illegal cultivation, sale, and use of cannabis. The main psychoactive component of the cannabis plant is D9‐tetrahydrocannabinol (THC). The introduction of mobile forensic labs has greatly increased the testing efficiency of psychotropic substances, but the traditional methods of analysis might seem to be cumbersome for the individual experimenting with the scene of seizure. The incorporation of portable analytical devices in forensic field analysis is the approach in the right direction for the testing of drugs. In this paper, we introduce a paper‐based microfluidic device for the onsite detection of marijuana. The device comprises the fast blue B reagent (FBB) as the main reagent. The µPAD was used for the successful detection of seized marijuana. The µPAD‐based detection was combined with a UV‐Vis spectroscopic confirmation. The computational modeling of the absorbance spectra gave further insights into the product formation during the reaction of THC and FBB. This novel method helps in a faster analysis of marijuana, at a considerably lower cost; which is a great boost to the forensic community and the law enforcement authority. [ABSTRACT FROM AUTHOR]

Iodine is an essential micronutrient for humans due to its fundamental role in the biosynthesis of thyroid hormones. As a key parameter to assess health conditions, iodine intake needs to be monitored to ascertain and prevent iodine deficiency. Iodine is available from various food sources (such as seaweed, fish, and seafood, among others) and dietary supplements (multivitamins or mineral supplements). In this work, a microfluidic paper-based analytical device (μPAD) to quantify iodide in seaweed and dietary supplements is described. The developed μPAD is a small microfluidic device that emerges as quite relevant in terms of its analytical capacity. The quantification of iodide is based on the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) by hydrogen peroxide in the presence of iodine, which acts as the catalyst to produce the blue form of TMB. Additionally, powder silica was used to intensify and uniformize the colour of the obtained product. Following optimization, the developed μPAD enabled iodide quantification within the range of 10–100 µM, with a detection limit of 3 µM, and was successfully applied to seaweeds and dietary supplements. The device represents a valuable tool for point-of-care analysis, can be used by untrained personnel at home, and is easily disposable, low-cost, and user-friendly.

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. [ABSTRACT FROM AUTHOR]

Here, we propose a microfluidic paper-based analytical device (µPAD) implemented with a near-field communication (NFC) tag as a portable, simple and fast colorimetric method for glutathione (GSH) determination. The proposed method was based on the fact that Ag+ could oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) into oxidized blue TMB. Thus, the presence of GSH could cause the reduction of oxidized TMB, which resulted in a blue color fading. Based on this finding, we developed a method for the colorimetric determination of GSH using a smartphone. A µPAD implemented with the NFC tag allowed the harvesting of energy from a smartphone to activate the LED that allows the capture of a photograph of the µPAD by the smartphone. The integration between electronic interfaces into the hardware of digital image capture served as a means for quantitation. Importantly, this new method shows a low detection limit of 1.0 µM. Therefore, the most important features of this non-enzymatic method are high sensitivity and a simple, fast, portable and low-cost determination of GSH in just 20 min using a colorimetric signal. [ABSTRACT FROM AUTHOR]

Multiplex detection can enhance diagnostic precision and improve diagnostic efficiency, providing important assistance for epidemiological investigation and epidemic prevention. There is a great need for multi-detection sensing platforms to accurately diagnose diseases. Herein, we reported a μPAD-based chemiluminescence (CL) assay for ultrasensitive multiplex detection of AIV biomarkers, based on three DNAzyme/Lum/PEI/CaCO 3. Three time-resolved CL signals were sequentially generated with detection limits of 0.32, 0.34, and 0.29 pM for H1N1, H7N9, and H5N1, respectively, and with excellent selectivity against interfering DNA. The recovery test in human serum displayed satisfactory analysis capabilities for complex biological samples. The μPAD-based CL assay achieved multiplex detection within 70 s, with a high time resolution of 20 s. The proposed strategy has the advantages of low cost, high sensitivity, good selectivity, and wide time resolution, the μPAD-based CL assay has shown great potential in the early and accurate diagnosis of diseases. We reported a μPAD-based CL assay for ultrasensitive multiplex detection of AIV biomarkers, based on three DNAzyme/Lum/PEI/CaCO 3. The platform achieved multiplex detection within 70 s, with a wide time resolution of 20 s, demonstrating great potential in the early and accurate diagnosis of diseases. [Display omitted] • A low-cost, portable μPAD with time-resolved CL signals was fabricated. • The CL assay successfully detected H1N1, H7N9, and H5N1 within 70 s, with a wide time resolution of 20 s. • Dual amplification of the sensor was achieved based on the CHA reaction and DNAzyme catalysis. • The CL assay had a wide linear range (1.0 pM−10 nM) and high detection sensitivity at pM level. • The CL assay was able to visualize trace H1N1, H7N9, and H5N1 down to 10 pM. [ABSTRACT FROM AUTHOR]

The rapid emergence of microfluidic paper-based devices as point-of-care testing (POCT) tools for early disease diagnosis and health monitoring, particularly in resource-limited areas, holds immense potential for enhancing healthcare accessibility. Leveraging the numerous advantages of paper, such as capillary-driven flow, porous structure, hydrophilic functional groups, biodegradability, cost-effectiveness, and flexibility, it has become a pivotal choice for microfluidic substrates. The repertoire of microfluidic paper-based devices includes one-dimensional lateral flow assays (1D LFAs), two-dimensional microfluidic paper-based analytical devices (2D μPADs), and three-dimensional (3D) μPADs. In this comprehensive review, we provide and examine crucial information related to paper substrates, design strategies, and detection methods in multi-dimensional microfluidic paper-based devices. We also investigate potential applications of microfluidic paper-based devices for detecting viruses, metabolites and hormones in non-invasive samples such as human saliva, sweat and urine. Additionally, we delve into capillary-driven flow alternative theoretical models of fluids within the paper to provide guidance. Finally, we critically examine the potential for future developments and address challenges for multi-dimensional microfluidic paper-based devices in advancing noninvasive early diagnosis and health monitoring. This article showcases their transformative impact on healthcare, paving the way for enhanced medical services worldwide. [Display omitted] • Structure design strategies for microfluidic paper-based analytical devices. • Sensing mechanism and application in noninvasive testing. • Theoretical models of capillary-driven fluids within paper. [ABSTRACT FROM AUTHOR]

A dual colorimetric/electrochemical sensor integrated with a microfluidic paper-based analytical device (μPAD) is created for rapid and sensitive detection of cabaryl in fruit. For colorimetric sensor, quercetin modified silver nanoparticle (QCT-AgNPs) is synthesized via self-reduction, resulting in the color change from green to dark green in the presence of cabaryl, which can be readily observed via naked eyes. This colorimetric sensor provides a linear response of 5 – 50 μM quantified by Image J software. For electrochemistry, AgNPs/MXene nanocomposite electrode offers high electrocatalytic activity towards the detection of carbarly with a linear range of 0 – 60 µM and a detection limit of 0.01 µM. The limit of detection values obtained from this dual sensor are lower than a cut-off of carbaryl residue in food, which is 7 mg.kg−1 (35 µM) verifying the desired sensitivity of this platform. Ultimately, this dual sensor is successfully applied for on-site detection of carbaryl in the orange samples read out on the smartphone with satisfactory results. [Display omitted] • Dual colorimetric/electrochemicl sensor is developed on μPAD for cabaryl sensor • QCT-AgNPs is synthesized by self-reduction and applied for colorimetric detection. • AgNPs/MXene modified electrode provided high electrochemical sensitivity. • This platform is applied for on-site carbaryl detection in fruits on smartphone. [ABSTRACT FROM AUTHOR]

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.

Here, we propose a microfluidic paper-based analytical device (µPAD) implemented with a near-field communication (NFC) tag as a portable, simple and fast colorimetric method for glutathione (GSH) determination. The proposed method was based on the fact that Ag+ could oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) into oxidized blue TMB. Thus, the presence of GSH could cause the reduction of oxidized TMB, which resulted in a blue color fading. Based on this finding, we developed a method for the colorimetric determination of GSH using a smartphone. A µPAD implemented with the NFC tag allowed the harvesting of energy from a smartphone to activate the LED that allows the capture of a photograph of the µPAD by the smartphone. The integration between electronic interfaces into the hardware of digital image capture served as a means for quantitation. Importantly, this new method shows a low detection limit of 1.0 µM. Therefore, the most important features of this non-enzymatic method are high sensitivity and a simple, fast, portable and low-cost determination of GSH in just 20 min using a colorimetric signal.

Nowadays, developing a cost-effective, easy-to-operate, and efficient signal amplification platform is of important to microfluidic paper-based analytical devices (μPAD) for end-use markets of point-of-care (POC) assay applications. Herein, an ultrasensitive, paper-based photoelectrochemical (PEC) bioassay platform is constructed by in situ grown ZnO/ZnIn2S4 heterojunctions onto paper fibers, which acted as photoactive signal amplification probes for enhancing the sensitivity of antibodies-based diagnostic assays, for the sensitive detection of alpha-fetoprotein (AFP) targets. The crystalline flake-like ZnIn2S4 composited with hexagonal nanorods (NRs) morphology of ZnO is an in situ grown, at the first time, onto cellulose fibers surface supported with Au nanoparticle (Au NP) modification to improve conductivity of the device working zone. The obtained composites on paper fibers are implemented as a flexible paper-based photoelectrode to realize remarkable performance of the fabricated μPAD, resulting from the enhanced PEC activity of heterojunctions with effective electron-hole pair separation for accelerating photoelectric conversion efficiency of the sensing process under light irradiation. Once the target AFP was introduced into the biosensing interface assistant, with a specific recognition interaction of AFP antibody, a drastically photocurrent response was generated, in view of the apparent steric effects. With the concentration increase of AFP targets, more immune conjugates could be confined onto the biosensing interface, eventually leading to the quantitative decrease of photocurrent intensity. Combined with an ingenious origami design and permitting the hydrophobic/hydrophilic conversion procedure in the bioassay process, the ultrasensitive PEC detection of AFP targets was realized. Under the optimized conditions, the level of AFP could be sensitively tracked by the prepared μPAD with a liner range from 0.1 to 100 ng mL−1 and limit of detection of 0.03 ng mL−1. This work provides a great potential application for highly selective and sensitive POC testing of AFP, and finally, developments for clinical disease diagnosis.

A microfluidic paper-based analytical device (μPAD) is developed in this work to analyze hydroquinone in facial whitening creams using phloroglucinol. The μPAD features a hydrophobic barrier for detection and was fabricated using a wax printer with Whatman chromatographic paper. Detection was achieved by colorimetry based on the formation of an orange hydroquinone-phloroglucinol complex. The colored reaction product formed on the detection zone of the μPAD was scanned, and the images obtained were processed with Image-J software to determine their color intensity (RGB value). Optimization of the process conditions was conducted to achieve sensitive measurements. The optimum conditions yielding maximum sensitivity included a reagent addition sequence of phloroglucinol → NaOH → sample (hydroquinone), 1 μL of 0.5% phloroglucinol, 1 M NaOH, and 10-minute reaction. Under optimal conditions, the μPAD produced two linear calibration curves for hydroquinone at concentrations of 10-100 mg/L (R² = 0.9979) and 250-1000 mg/L (R² = 0.9991). The method demonstrated very good selectivity for the target analyte in the presence of propylene glycol and resorcinol with satisfactory validity and average recovery close to 100%. The proposed μPAD is a very simple and inexpensive technique for hydroquinone analysis and could be applied to cosmetics samples with satisfactory results. [ABSTRACT FROM AUTHOR]

[Display omitted] • Paper-based analytical aptasensor for food safety and quality. • Determination of β-lactoglobulin traces in food products. • Aptamer bioconjugation to fluorescent carbon quantum dots. • Smartphone-assisted sensing platform for portable β-lactoglobulin determination. Herein, we present a new luminescent-based analytical method for sensitive fluorometric determination of a milk allergen, β-lactoglobulin (β-LG), in food products based on a portable, low-cost, and easy-to-use microfluidic paper-based analytical device (µPAD). For this purpose, the developed µPAD consisted on a β-LG specific aptamer immobilized on a cellulose disk, and fluorescent Carbon Quantum Dots (CQDs) bioconjugated with its complementary oligonucleotide (DNA), which worked as a label for the aptamer hybridization and its complementary DNA (CQDs-DNA). In a sample with β-LG, the aptamer preferentially binds to β-LG protein and this leads to release of the CQDs-DNA. After washing the µPAD, where the aptamer is immobilized, detection is achieved by fluorescence intensity decrease. An estimation of β-LG concentration can be calculated thanks to the fluorescence changes, for that, a photo of the µPAD is taken using either a digital camera or a smartphone. The µPAD showed an excellent detection response for β-LG in the range of 1.8 to 500 ng/mL with a limit of detection of up to 0.6 ng/mL using the digital camera. Furthermore, the developed method was successfully applied to the determination of β-LG in food samples such as skimmed milk, crispy rice, cookies and chocolate. [ABSTRACT FROM AUTHOR]

Hydrogen peroxide has been postulated to be present on the surface of Europa and Enceladus. While it could represent a potential source of energy for possible life-forms, H2O2 may also interfere with a number of current detection technologies, including biosensors. To take advantage of the selectivity and portability of these devices, simple and reliable routes to degrade the potential H2O2 present should be developed and implemented to prepare for this possibility. Unfortunately, most of the current approaches for removing H2O2 are slow, may affect the sample, or could interfere with the performance of biosensors. To address these limitations, catalase was immobilized onto silica particles and used as a means to selectively decompose H2O2 prior to the analysis of common biomarkers with a biosensor. For these experiments, glucose, l-leucine, and lactic acid were used as representative examples of biomolecules such as carbohydrates, amino acids, and organic acids, respectively, which could be used as biomarkers on extraterrestrial bodies. While the decomposition reaction between catalase and H2O2 is well known, to our knowledge this is the first instance where catalase has been used in combination with a microfluidic paper-based analytical device (μPAD) to implement selective sample pretreatment. [ABSTRACT FROM AUTHOR]

Despite recent advancements in the field of microfluidic paper-based analytical devices (μPADs), a key challenge remains in developing a simple and efficient μPAD with customized imaging capabilities for antioxidant assays. In the present study, we report a facile approach for μPAD fabrication through the application of transparent nail paint leading to creation of hydrophobic barriers and well-defined channels. The resultant μPADs were then characterized through scanning electron microscopy and contact angle measurements. The resolution and functional features of the fabricated μPAD were amenable to the intended assay. The μPAD's impregnated poly(methacrylic acid) (PMAA)-coated cerium oxide (CeO2) nanoparticles oxidized the 3,3′,5,5′-tetramethylbenzidine (TMB) leading to the formation of a blue-colored charge-transfer complex. The addition of different antioxidant standard solutions resulted in a reduction in the blue color in a dose-dependent manner which could be observed visually. The color intensity of the PMAA-CeO2 nanoparticle@TMB oxidation product was inversely proportional to the antioxidant concentration and was measured using customized in-house MATLAB-based image processing software. Importantly, PMAA-CeO2 nanoparticle-based μPADs demonstrated good analytical characteristics and were able to be stored for long periods without any loss of activity. Moreover, potential interferents did not pose any threat to the colorimetric signal read-out for determination of antioxidant activity. The developed method was further applied for the assessment of antioxidant activity in a variety of tea samples and performed satisfactorily in comparison with a commonly used antioxidant detection method. Collectively, the developed μPAD-based platform holds great potential as a low-cost, convenient, portable and reliable method for pursuing various on-site antioxidant assays. [ABSTRACT FROM AUTHOR]

In human history, the paper has long been used as a platform to record and preserve information. However, over the decades, paper has found its application in Biomedical Sciences, too. Both paper-based microfluidic devices (μPADs) and paper-based cultures and scaffolds have shown immense potential to be used as a sensor as well as a supporting material for in vitro tissue engineering. μPADs can be used to perform low-cost and fast biomolecular assays at Point-Of-Care (POC). They are being used to detect various biomarkers like viral proteins, metabolites, oncogenes, and antigens; and conditions like Venous Thromboembolism (VTE). On the other hand, the paper has also been used to develop paper-based 3D cultures and scaffolds to test drugs, and monitor cytotoxic effects in vitro cell microenvironments and also as implantable tissues. In this review, we intend to enumerate the development in the field of μPADs, paper-based cell cultures, and paper-based scaffolds and their plethora of applications over the last decade.

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 MgFe 2 O 4 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. [Display omitted] • A simple electrochemical microfluidic paper-based device • Non-enzymatic detection of Ascorbic Acid (AA) • A microfluidic device for blood-plasma separation with high efficacy • AA detection in miniaturized POC setting via single step [ABSTRACT FROM AUTHOR]

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. [Display omitted] • A 3D paper-based microfluidic device (μPAD) was designed via origami methods. • Sliding paper tape as a valve for fluid control. • μPAD successfully applied to the detection of Fe (III), Ni (II), Cr (VI) and nitrite. • 3D μPAD is a convenient and efficient rapid detection device. [ABSTRACT FROM AUTHOR]

The medical field and public health rely on diagnostics carried out by sensors and laboratory testing methods. However, conducting laboratory tests is a demanding task, requiring both substantial equipment and the skilled touch of experienced operators. The microfluidic field has tackled this challenge over recent years. Paper microfluidic devices have become popular recently due to their testing speed, low cost, and accessibility. They are more suitable for field operations and developing regions with limited access to centralized healthcare centers and medical laboratories since they do not require external flow control instruments and are much cheaper to make when compared to conventional microfluidic chips. However, the majority of paper-based analytical devices tend to be qualitative or semi-quantitative; consequently, there is a significant need for paper-based quantitative transduction methods, given their expanding range of applications. Here, we introduce a novel microwave paper-based metamaterial-inspired transduction method that relies on dielectric sensing. Using paper as both the microfluidic channel and the resonator substrate hugely reduces the sensing device's costs, complexity, and environmental impact. We designed the metamaterial-based transducer using computational methods and demonstrated its fabrication and operation experimentally. Dielectric sensing of multiple analytes with different permittivities is demonstrated as a proof of concept. Although the device has a non-linear response curve in a wide range of permittivity change, the response can be assumed linear in the shorter spans. The results for permittivities lower than 30 show a 2.14 MHz/RPU sensitivity. This design is a prototype demonstrating the possibility of integrating a porous media microfluidic channel and paper-based microwave resonators. The linear sensing performance illustrated by the platform indicates its potential applications in the biosensing field. [Display omitted] • A novel paper-based metamaterial-inspired transducer is introduced. • Paper serves as microfluidic channel and substrate, reducing costs and eco impact. • This unique transduction method is a new type of quantitative analysis for μ PADs. • The linear sensing concept is demonstrated using various solutions. • This approach has implications for affordable quantitative μ PADs. [ABSTRACT FROM AUTHOR]

Microfluidic paper combines pump-free water transport at low cost with a high degree of sustainability, as well as good availability of the paper-forming cellulosic material, thus making it an attractive candidate for point-of-care (POC) analytics and diagnostics. Although a number of interesting demonstrators for such paper devices have been reported to date, a number of challenges still exist, which limit a successful transfer into marketable applications. A strong limitation in this respect is the (unspecific) adsorption of protein analytes to the paper fibers during the lateral flow assay. This interaction may significantly reduce the amount of analyte that reaches the detection zone of the microfluidic paper-based analytical device (µPAD), thereby reducing its overall sensitivity. Here, we introduce a novel approach on reducing the nonspecific adsorption of proteins to lab-made paper sheets for the use in µPADs. To this, cotton linter fibers in lab-formed additive-free paper sheets are modified with a surrounding thin hydrogel layer generated from photo-crosslinked, benzophenone functionalized copolymers based on poly-(oligo-ethylene glycol methacrylate) (POEGMA) and poly-dimethyl acrylamide (PDMAA). This, as we show in tests similar to lateral flow assays, significantly reduces unspecific binding of model proteins. Furthermore, by evaporating the transport fluid during the microfluidic run at the end of the paper strip through local heating, model proteins can almost quantitatively be accumulated in that zone. The possibility of complete, almost quantitative protein transport in a µPAD opens up new opportunities to significantly improve the signal-to-noise (S/N) ratio of paper-based lateral flow assays.

Paper is an ideal candidate for the development of new disposable diagnostic devices because it is a low-cost material, allows transport of the liquid on the device by capillary action, and is environmentally friendly. Today, colorimetric analysis is most often used as a detection method for rapid tests (test strips or lateral flow devices) but usually gives only qualitative results and is limited by a relatively high detection threshold. Here, we describe studies using fluorescence as a readout tool for paper-based diagnostics. We study how the optical readout is affected by light transmission, scattering, and fluorescence as a function of paper characteristics such as thickness (grammage), water content, autofluorescence, and paper type/composition. We show that paper-based fluorescence analysis allows better optical readout compared to that of nitrocellulose, which is currently the material of choice in colorimetric assays. To reduce the loss of analyte molecules (e.g., proteins) due to adsorption to the paper surface, we coat the paper fibers with a protein-repellent hydrogel. For this purpose, we use hydrophilic copolymers consisting of N , N -dimethyl acrylamide and a benzophenone-based cross-linker, which are photochemically transformed into a fiber-attached polymer hydrogel on the paper fiber surfaces in situ. We show that the combination of fluorescence detection and the use of a protein-repellent coating enables sensitive paper-based analysis. Finally, the success of the strategy is demonstrated by using a simple LFD application as an example.

• A μPAD with two layers was developed to monitor Cu2+, Fe2+ and free chlorine simultaneously. • The detection limits of μPAD for single analyte were less or equal to each Chinese Drinking Water Standard concentration. • The μPAD exhibited good applicability for real use in different kinds of drinking water. Cu2+, Fe2+ and free chlorine are three important analytes in drinking water. Firstly, the μPAD for single analyte was easily fabricated. Detection limits for Cu2+, Fe2+ and free chlorine were 0.1 mg L−1, 0.1 mg L−1 and 0.2 mg L−1, respectively, less or equal to each Chinese Drinking Water Standard concentration and lower than those of most commercially available testing strips. Another μPAD with two layers was then developed to monitor and report the safety level of these three analytes in drinking water simultaneously. Moreover, the μPADs for Fe2+ and free chlorine provided more distinguishable colors to users with naked eyes. The as-fabricated μPAD not only demonstrated excellent anti-interference capability, production reproducibility, but also exhibited good applicability for real use in different kinds of drinking water. [ABSTRACT FROM AUTHOR]

Abstract The design, optimization and evaluation of a micro paper-based analytical device (μPAD) for the simultaneous determination of fluoride and nitrite in real water samples was developed. In order to create hydrophilic and hydrophobic zones on the laboratory filter paper device, a wax ink printer was used. The μPAD has four reaction zones for each analyte where the respective colorimetric reactions took place using the SPADNS method and Griess method for fluoride and nitrite respectively. A Smartphone was used to capture the corresponding images which were then converted to the RGB colour space by the software ImageJ using a personal computer. Under the optimal conditions, the linear range was 0.23–2.26 mg L−1 for fluoride with limit of detection (LOD) and limit of quantification (LOQ) of 0.13 mg L−1 and 0.25 mg L−1. The working range for nitrite was 0.05–10.0 mg L−1 with LOD of 0.03 mg L−1 and LOQ 0.13 mg L−1. The μPAD was used to determine the analytes in water samples obtaining satisfactory results in both cases. The validation of fluoride determination was performed comparing the μPAD with the reference method and for nitrite determination, a recovery study was carried out. In both cases, the obtained results demonstrated a good agreement. Graphical abstract Unlabelled Image Highlights • The design, optimization and evaluation of a micro paper-based analytical device was developed. • This user-friendly μPAD simultaneously determines fluoride and nitrite in real water samples. • A commercial wax printer was employed to create this portable inexpensive device on filter paper. • It requires minimum volumes of sample and reagents and uses biodegradable raw materials. • Results demonstrate this device is a promising analytical tool for a wide field of applications. [ABSTRACT FROM AUTHOR]

The main problem for the expansion of the use of microfluidic paper-based analytical devices and, thus, their mass production is their inherent lack of fluid flow control due to its uncontrolled fabrication protocols. To address this issue, the first step is the generation of uniform and reliable microfluidic channels. The most common paper microfluidic fabrication method is wax printing, which consists of two parts, printing and heating, where heating is a critical step for the fabrication of reproducible device dimensions. In order to bring paper-based devices to success, it is essential to optimize the fabrication process in order to always get a reproducible device. Therefore, the optimization of the heating process and the analysis of the parameters that could affect the final dimensions of the device, such as its shape, the width of the wax barrier and the internal area of the device, were performed. Moreover, we present a method to predict reproducible devices with controlled working areas in a simple manner.

Acid dissociation constant is an important chemical characteristic of organic and inorganic compounds and it affects both chemical properties and biological activities of the molecules. Herein, a very simple, fast and cost-effective method based on microfluidic technology has been reported for colorimetric determination of acidity constants. The designed device works based on pH-metric titration of colorful indicators followed by colorimetric measurements by a smart phone mobile device or a flatbed scanner. So, it does not need sophisticated instrumentation and is accomplished in a very short time (about 1 min). All titration steps are transferred on a star-like designed µPAD device: (1) spotting 0.3 µL buffers of different pHs at the end of the channels (reaction zones), (2) spotting a 30 µL portion of the indicator on the center of µPAD followed by movement of indicator solution toward the reaction zone by capillary action of the paper. The measured color change of the indicators at the reaction zone is fitted to the Henderson-Hasselbalch equation, through which acidity constants are calculated. The performance of the device was evaluated by measuring acidity constant of 4 indicators including bromothymol blue, bromocresol green, bromocresol purple and phenolphthalein. A very close agreement was achieved between those measured by the suggested device here and the previously reported values. The reproducibility of this method was lower than 5% for relative standard deviation of three replicate measurements. [ABSTRACT FROM AUTHOR]

The utility of strikingly simple and cheap optoelectronic detectors operating according to a Paired-Emitter-Detector-Diode (PEDD) principle for microfluidic paper-based analytical devices (μPAD) has been experimentally confirmed. The prototypes of these compact detectors have been made of only two customary light emitting diodes without any additional optical parts like lens, filters, fibers etc. Moreover, for their operation economic and portable equipment is required (low-power circuit and ordinary voltmeter). Photometric and fluorometric PEDDs for paper devices are developed. In both cases the possibility of their applicability in transmittance and reflectance modes of measurements are presented. In these investigations as model analytes some coloured and fluorescence substances have been handled, but the utility of several developed systems for real chemical analysis has also been demonstrated. The results of photometric determination of hemoglobin in human blood, as well as fluorometric determination of quinine in tonics and calcium ions in mineral waters using various PEDD-μPAD systems have been shown. [ABSTRACT FROM AUTHOR]

A microfluidic paper-based analytical device (μPADs) immunoassay for detection of the blood native biomarker D-dimer is reported. The μPAD is created by wax printing on a single piece of chromatographic paper and combined with an anti-D-dimer capture antibody and conjugates of anti-D-dimer antibody with 40 nm gold nanoparticles. The presence of D-dimer in buffer/simulated plasma samples is successfully reported for concentrations as low as 15 ng D-dimer/mL. Linearity between signal intensity and D-dimer concentration is observed up to 100 ng/mL. Using an appropriate dilution, the test could be used to yield positive results only for those samples with a D-dimer concentration above the clinically relevant threshold range of 250–500 ng/mL. Finally, the merits and pitfalls of using μPADs as compared to lateral flow devices in immunoassays are discussed. [ABSTRACT FROM AUTHOR]

[Display omitted] • Iron speciation of Iron(II) and Iron(III) in wines using a handheld-size microfluidic paper-based device. • Single device assembled for quantification of both ionic forms accounting for sample colour interference. • Application to both white and red wines in a portable, in situ, approach. During the winemaking process, wine is frequently exposed to oxygen which can affect the organoleptic characteristics and therefore the quality of the finished wine. Most phenolic molecules in wine serve as oxidation substrates affecting colour and flavour. Transition metal ions namely copper and iron act as electron pumps; in fact, Fe(III) is reduced to Fe(II) as a consequence of the oxidation of phenols to quinones and, in the presence of oxygen, Fe(II) is oxidised back to Fe(III). Consequently, speciation analysis by the oxidation state is critical and even more relevant than the total ion concentration when one wants to evaluate the risk for wine oxidation. Interestingly the different reactivity of Fe(III) and Fe(II) makes spectrophotometric methods, namely colorimetric reactions, ideally suited for speciation analysis. In this work, a microfluidic paper-based analytical device (µPAD) was designed for the iron speciation in wine samples. The colorimetric reaction between bathophenanthroline and iron(II) was chosen and the oxidation of Fe(III) to Fe(II), using hydroxylamine, was carried out within the same device. Aiming for application to different types of wine, the developed device also enabled to account for the potential sample colour (subtraction of interfering signal). The designed µPAD enabled the iron speciation in both white and red wines. [ABSTRACT FROM AUTHOR]

The following article summarizes United States Patent Application No. US20180052155A1, titled ‘Assay Devices and Methods’ (filed 16 August 2016, published 22 February 2018). While lateral flow assays (LFAs) have revolutionized point-of-care diagnostics by enabling accurate, inexpensive, and rapid detection of biomarkers, they typically do not provide quantitative results. Hence, there is a significant need for quantitative assays at the point of care. This patent summary describes a novel method of chronometric biomarker quantitation via enzymatic degradation of a metastable gelatin-based biomatrix, principally suited for use in paper-based microfluidic devices (microPADs). This new quantitation mechanism was designed to meet the ASSURED criteria for point-of-care diagnostic devices laid forth by the World Health Organization and may ultimately provide increased access to healthcare, at a significantly reduced cost, around the world.

Paper-based microfluidic devices have advanced significantly in recent years as they are affordable, automated with capillary action, portable, and biodegradable diagnostic platforms for a variety of health, environmental, and food quality applications. In terms of commercialization, however, paper-based microfluidics still have to overcome significant challenges to become an authentic point-of-care testing format with the advanced capabilities of analyte purification, multiplex analysis, quantification, and detection with high sensitivity and selectivity. Moreover, fluid flow manipulation for multistep integration, which involves valving and flow velocity control, is also a critical parameter to achieve high-performance devices. Considering these limitations, the aim of this review is to (i) comprehensively analyze the fabrication techniques of microfluidic paper-based analytical devices, (ii) provide a theoretical background and various methods for fluid flow manipulation, and (iii) highlight the recent detection techniques developed for various applications, including their advantages and disadvantages.

Microfluidic paper-based analytical devices (μPADs) have a significant potential in developing portable and disposable point-of-care testing (POCT). Herein, a facile, rapid, cost-effective and environment friendly strategy for μPADs fabrication is proposed. Specifically, the substrate paper was hydrophobized by coating with trimethoxysilane (TOS), and then the selected area was hydrophilized by treating with surfactant. The whole fabrication process was implemented within 7 min, with no need for complex pre-treatment, high-temperature and special equipment. As a proof-of-concept application, the as-prepared μPAD was applied to determination of the glucose content in human serum samples. The results agreed well with those obtained by a glucometer. We believe that the μPADs fabrication method proposed here could provide a facile, rapid and low-cost reference for other related studies. [ABSTRACT FROM AUTHOR]

A microfluidic biosensor paper-based analytical device, or μPAD, was developed as a point-of-care assay platform for the simultaneous determination of three clinically important markers: uric acid, urea, and triglycerides. To achieve a simultaneous determination, thermosensitive valves based on N-isopropylacrylamide polymers controlled the flow of fluid in the μPAD. The evaluation of the analytical characteristics of the μPAD was done using a photographic camera at room temperature. The detection limits were 4.5·10−5, 2.5·10−3, and 1.5·10−3 mg mL−1, for uric acid, urea, and triglycerides, respectively, and the precision expressed as relative standard deviation (% RSD) 3.2, 1.2, and 1.6%. The obtained μPAD devices were validated with real human plasma samples, demonstrating high accuracy and precision. Multiple analyte point-of-care tests such as those developed here have excellent characteristics, being easy to manufacture, cost-effective, easy to use, and highly sensitive. [Display omitted] • A point-of-care assay platform based μPAD device was developed for the simultaneous determination of three clinically important markers: uric acid, urea, and triglycerides. • Thermosensitive valves based on isopolyacrylamide-acrylamide hydrogels controlled the flow of fluid in the μPAD. • The obtained μPAD devices were validated with real human plasma samples, demonstrating high accuracy and precision. • μPAD device has excellent characteristics: easy to manufacture and to use, cost-effective and high sensitivity. [ABSTRACT FROM AUTHOR]

The synthesis of a novel spirooxazine (1-benzyl-3,3-dimethylspiro[indoline-2,3′-naphtho [2,1-b] [1,4] oxazine, BSP-SP) was developed as a colorimetric probe for the quantitative analysis of Fe2+ and Pb2+ in water samples. The device is based on a microfluidic paper-based analytical device (μPAD). BSP-SP was characterized by various techniques, e.g. single-crystal X-ray diffraction (SC-XRD) and time-dependent density functional theory (TD-DFT). In addition, the density functional theory (DFT) allowed us to understand better the adsorption of Fe2+ and Pb2+ over other cations adsorption on the BSP-MO structure. The structural isomerization of BSP-SP occurs as an open-form merooxazine (BSP-MO), which acts as a ligand to form Fe2+/BSP-MO and Pb2+/BSP-MO complexes. We observed these complexes under UV irradiation as having a light blue coloration. The μPADs were also proposed as detection platforms to maintain photostability and photochromic efficiency and extend the lifetime of the spirooxazine. Under optimal conditions, the developed μPAD enables rapid and sensitive quantitative analysis of Fe2+ at pH 6 and Pb2+ at pH 11. The limits of detection (LOD) were 0.036 mg L−1 for Fe2+ and 0.152 mg L−1 Pb2+. Overall, the developed sensors, particularly the μPADs, promise the practical analysis of Fe2+ and Pb2+ in water samples. [Display omitted] • Our novel spirooxazine (BSP) was successfully synthesized as colorimetric probe. • BSP-coated μPAD was illustrated to enhance photochromic efficiency. • This method features selectivity toward Fe2+ and Pb2+ determination. [ABSTRACT FROM AUTHOR]

This work demonstrates the fabrication of microfluidic paper-based analytical devices (°PADs) suitable for the analysis of sub-microliter sample volumes. The wax-printing approach widely used for the patterning of paper substrates has been adapted to obtain high-resolution microfluidic structures patterned in filter paper. This has been achieved by replacing the hot plate heating method conventionally used to melt printed wax features into paper by simple hot lamination. This patterning technique, in combination with the consideration of device geometry and the influence of cellulose fiber direction in filter paper, led to a model PAD design with four microfluidic channels that can be filled with as low as 0.5 μL of liquid. Finally, the application to a colorimetric model assay targeting total protein concentrations is shown. Calibration curves for human serum albumin (HSA) were recorded from sub-microliter samples (0.8 μL), with tolerance against ±0.1 μL variations in the applied liquid volume. [ABSTRACT FROM AUTHOR]

Paper based microfluidics (µPADs) with advantages of portability, low cost, and ease of use have attracted extensive attention. Here we describe a novel method that integrates glucoamylase-trapped aptamer-crosslinked hydrogel for molecular recognition with cascaded enzymatic reactions for signal amplification and a µPAD for portable readout. Upon target introduction, the hydrogel decomposes to release glucoamylase, which catalyzes the hydrolysis of amylose to produce a large amount of glucose. With a simple folding of the µPAD, the sample solution containing glucose product wicks and diffuses in parallel to each test-zone to carry out homogeneous assays, where glucose is used to produce I 2 for brown color visualization through multiple enzymatic and chemical cascade reactions. Through color gradient changes based on different concentrations of the target, a semiquantitative assay is achieved by the naked eye, and quantitation can be obtained by handheld devices. Detection of cocaine in buffer and urine was performed to demonstrate the utility of the hydrogel–µPAD system. More importantly, the hydrogel–µPAD system can be extended to the detection of various targets by incorporating the corresponding aptamer into the hydrogel. The hydrogel–µPAD system reported here provides a new platform for portable, disposable and visual detection of a wide range of targets. [ABSTRACT FROM AUTHOR]

[Display omitted] • ABO and Rh blood type detection using a three-dimensional origami microfluidic technology. • Blood moves through paper thickness instead of paper length. • The device simply works based on the counting the number of layers through which blood moves. • The proposed method can determine the blood type of 95% of samples compared to the gold method. We report a three-dimensional origami paper-based analytical device (3D-µPAD) for ABO and Rh blood type detection using a three-dimensional origami microfluidic technology. A piece of rectangular shaped paper sizing (28.5 × 2) cm was folded into thirteen square layers of 2 cm edge. The first two layers formed the sample zone and distribution zones. Other layers were composed of 4 hydrophilic circle zones, which provide cylindrical channels when the paper layers are folded. Three circle zones on layers from 3 to 8 were pre-deposited with antibodies A, B and D and the reminder circle zone was considered as control. According to the design principle, when blood moves through channels of different antibodies it forms agglutination with those having its complementary blood group antigen. This lowers down the speed of blood movement through the channels. Therefore, the ability of blood movement through each channel was used as the basis of our detection (i.e., counting the number of layers through which blood moved for each channel, separately). The effects of the experimental parameters on the sensor's performance were investigated by using Placket-Burman and Central Composite Design methods. The device operation was validated by analyzing 67 blood samples. It was found that the if blood had complementary antigens, it moved 3–5 layers in the channels pre-deposited with anti-A or anti-B and 5–7 layer in channels pre-deposited with anti-D. Based on this observation, by single analysis we could determine the blood type of 95 % of samples compared to the gold standard method, which is currently used in blood transfusion labs. Replicate analysis resulted in 100 % accuracy. The device was stable for 14 days before using. However, after using, it was stable for at least 1 year. [ABSTRACT FROM AUTHOR]

Zika virus (ZIKV) infection may cause serious birth defects and is a critical concern for women of child-bearing age in affected regions. A simple, portable, and easy-to-use ZIKV detection method would enable point-of-care testing, which may aid in prevention of the spread of the virus. Herein, we describe a reverse transcription isothermal loop-mediated amplification (RT-LAMP) method that detects the presence of ZIKV RNA in complex samples (e.g., blood, urine, and tap water). Phenol red is the colorimetric indicator of successful amplification. Color changes based on the amplified RT-LAMP product from the presence of viral target are monitored using a smartphone camera under ambient light conditions. A single viral RNA molecule per μL can be detected in as quickly as 15 min using this method with 100% sensitivity and 100% specificity in blood and tap water, while 100% sensitivity and 67% specificity in urine. This platform can also be used to identify other viruses including SARS-CoV-2 and improve the current state of field-based diagnostics., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Paper-based systems have been widely investigated for developing point-of-care devices because of their simplicity, affordability, and ease of use. Recent advances have resulted in paper systems that have progressed beyond the historical “single-strip” format and allow for a larger range of functions. This review provides a summary of the advances that have been made to improve the utility of paper-based diagnostic tests for biosensing. Specifically, techniques for designing paper devices, including different geometries and chemical patterning to control fluid flow, are discussed. This review also examines novel approaches to improve paper-based assay sensitivities, such as sample preconcentration, signal amplification at the detection zone, and electrochemical methods. [ABSTRACT FROM AUTHOR]