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This study presents a portable, low-cost, point-of-care (POC) system for the simultaneous detection of blood glucose and hematocrit. The system consists of a disposable origami microfluidic paper-based analytical device (μPAD) for plasma separation, filtration, and reaction functions and a 3D-printed cassette for hematocrit and blood glucose detection using a smartphone. The origami μPAD is patterned using a cost-effective label printing technique instead of the conventional wax printing method. The 3D-printed cassette incorporates an array of LED lights, which mitigates the effects of intensity variations in the ambient light and hence improves the accuracy of the blood glucose and hematocrit concentration measurements. The hematocrit concentration is determined quantitatively by measuring the distance of plasma wicking along the upper layer of the origami μPAD, which is pretreated with sodium chloride and Tween 20 to induce dehydration and aggregation of the red blood cells. The filtered plasma also penetrates to the lower layer of the origami μPAD, where it reacts with embedded colorimetric assay reagents to produce a yellowish-brown complex. A color image of the reaction complex is captured using a smartphone inserted into the 3D-printed cassette. The image is analyzed using self-written RGB software to quantify the blood glucose concentration. The calibration results indicate that the proposed detection platform provides an accurate assessment of the blood glucose level over the range of 45-630 mg/dL (R 2 = 0.9958). The practical feasibility of the proposed platform is demonstrated by measuring the blood glucose and hematocrit concentrations in 13 human whole blood samples. Taking the measurements obtained from commercial glucose and hematocrit meters as a benchmark, the proposed system has a differential of no more than 6.4% for blood glucose detection and 9.1% for hematocrit detection. Overall, the results confirm that the proposed μPAD is a promising solution for cost-effective and reliable POC health monitoring.

Nanorod structures exhibit a high surface-to-volume ratio, enhancing the accessibility of electrolyte ions to the electrode surface and providing an abundance of active sites for improved electrochemical sensing performance. In this study, tetragonal α-MnO 2 with a large K + -embedded tunnel structure, directly grown on microfibrous carbon paper to form densely packed nanorod arrays, is investigated as an electrocatalytic material for non-enzymatic glucose sensing. The MnO 2 nanorods electrode demonstrates outstanding catalytic activity for glucose oxidation, showcasing a high sensitivity of 143.82 µA cm -2 mM -1 within the linear range from 0.01 to 15 mM, with a limit of detection (LOD) of 0.282 mM specifically for glucose molecules. Importantly, the MnO 2 nanorods electrode exhibits excellent selectivity towards glucose over ascorbic acid and uric acid, which is crucial for accurate glucose detection in complex samples. For comparison, a gold electrode shows a lower sensitivity of 52.48 µA cm -2 mM -1 within a linear range from 1 to 10 mM. These findings underscore the superior performance of the MnO 2 nanorods electrode in both sensitivity and selectivity, offering significant potential for advancing electrochemical sensors and bioanalytical techniques for glucose monitoring in physiological and clinical settings.

Specialized cancer treatments have the potential to exploit glutamine dependence to increase patient survival rates. Glutamine diagnostics capable of tracking a patient's response to treatment would enable a personalized treatment dosage to optimize the tradeoff between treatment success and dangerous side effects. Current clinical glutamine testing requires sophisticated and expensive lab-based tests, which are not broadly available on a frequent, individualized basis. To address the need for a low-cost, portable glutamine diagnostic, this work engineers a cell-free glutamine biosensor to overcome assay background and signal-to-noise limitations evident in previously reported studies. The findings from this work culminate in the development of a shelf-stable, paper-based, colorimetric glutamine test with a high signal strength and a high signal-to-background ratio for dramatically improved signal resolution. While the engineered glutamine test is important progress towards improving the management of cancer and other health conditions, this work also expands the assay development field of the promising cell-free biosensing platform, which can facilitate the low-cost detection of a broad variety of target molecules with high clinical value.

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.

A simple, equipment-free, direct fluorometric method, employing paper-based analytical devices (PADs) as sensors, for the selective determination of quinine (QN) is described herein. The suggested analytical method exploits the fluorescence emission of QN without any chemical reaction after the appropriate pH adjustment with nitric acid, at room temperature, on the surface of a paper device with the application of a UV lamp at 365 nm. The devices crafted had a low cost and were manufactured with chromatographic paper and wax barriers, and the analytical protocol followed was extremely easy for the analyst and required no laboratory instrumentation. According to the methodology, the user must place the sample on the detection area of the paper and read with a smartphone the fluorescence emitted by the QN molecules. Many chemical parameters were optimized, and a study of interfering ions present in soft drink samples was carried out. Additionally, the chemical stability of these paper devices was considered in various maintenance conditions with good results. The detection limit calculated as 3.3 S/N was 3.6 mg L -1 , and the precision of the method was satisfactory, being from 3.1% (intra-day) to 8.8% (inter-day). Soft drink samples were successfully analyzed and compared with a fluorescence method.

Vibration sensors are widely used in many fields like industry, agriculture, military, medicine, environment, etc. However, due to the speedy upgrading, most sensors composed of rigid or even toxic materials cause pollution to the environment and give rise to an increased amount of electronic waste. To meet the requirement of green electronics, biodegradable materials are advocated to be used to develop vibration sensors. Herein, a vibration sensor is reported based on a strategy of pencil-drawing graphite on paper. Specifically, a repeated pencil-drawing process is carried out on paper with a zigzag-shaped framework and parallel microgrooves, to form a graphite coating, thus serving as a functional conductive layer for electromechanical signal conversion. To enhance the sensor's sensitivity to vibration, a mass is loaded in the center of the paper, so that higher oscillation amplitude could happen under vibrational excitation. In so doing, the paper-based sensor can respond to vibrations with a wide frequency range from 5 Hz to 1 kHz, and vibrations with a maximum acceleration of 10 g. The results demonstrate that the sensor can not only be utilized for monitoring vibrations generated by the knuckle-knocking of plastic plates or objects falling down but also can be used to detect vibration in areas such as the shield cut head to assess the working conditions of machinery. The paper-based MEMS vibration sensor exhibits merits like easy fabrication, low cost, and being environmentally friendly, which indicates its great application potential in vibration monitoring fields.

As power sources for Internet-of-Things sensors, thermoelectric generators must exhibit compactness, flexibility, and low manufacturing costs. Stretchable and flexible painted thermoelectric generators were fabricated on Japanese paper using inks with dispersed p- and n-type single-walled carbon nanotubes (SWCNTs). The p- and n-type SWCNT inks were dispersed using the anionic surfactant of sodium dodecylbenzene sulfonate and the cationic surfactant of dimethyldioctadecylammonium chloride, respectively. The bundle diameters of the p- and n-type SWCNT layers painted on Japanese paper differed significantly; however, the crystallinities of both types of layers were almost the same. The thermoelectric properties of both types of layers exhibited mostly the same values at 30 °C; however, the properties, particularly the electrical conductivity, of the n-type layer increased linearly, and of the p-type layer decreased as the temperature increased. The p- and n-type SWCNT inks were used to paint striped patterns on Japanese paper. By folding at the boundaries of the patterns, painted generators can shrink and expand, even on curved surfaces. The painted generator (length: 145 mm, height: 13 mm) exhibited an output voltage of 10.4 mV and a maximum power of 0.21 μW with a temperature difference of 64 K at 120 °C on the hot side.

In the realm of electrochemical nitrite detection, the potent oxidizing nature of nitrite typically necessitates operation at high detection potentials. However, this study introduces a novel approach to address this challenge by developing a highly sensitive electrochemical sensor with a low reduction detection potential. Specifically, a copper metal nanosheet/carbon paper sensitive electrode (Cu/CP) was fabricated using a one-step electrodeposition method, leveraging the catalytic reduction properties of copper's high occupancy d-orbital. The Cu/CP sensor exhibited remarkable performance in nitrite detection, featuring a low detection potential of -0.05 V vs. Hg/HgO, a wide linear range of 10~1000 μM, an impressive detection limit of 0.079 μM (S/N = 3), and a high sensitivity of 2140 μA mM -1 cm -2 . These findings underscore the efficacy of electrochemical nitrite detection through catalytic reduction as a means to reduce the operational voltage of the sensor. By showcasing the successful implementation of this strategy, this work sets a valuable precedent for the advancement of electrochemical low-potential nitrite detection methodologies.

Image compression is a vital component for domains in which the computational resources are usually scarce such as automotive or telemedicine fields. Also, when discussing real-time systems, the large amount of data that must flow through the system can represent a bottleneck. Therefore, the storage of images, alongside the compression, transmission, and decompression procedures, becomes vital. In recent years, many compression techniques that only preserve the quality of the region of interest of an image have been developed, the other parts being either discarded or compressed with major quality loss. This paper proposes a study of relevant papers from the last decade which are focused on the selection of a region of interest of an image and on the compression techniques that can be applied to that area. To better highlight the novelty of the hybrid methods, classical state-of-the-art approaches are also analyzed. The current work will provide an overview of classical and hybrid compression methods alongside a categorization based on compression ratio and other quality factors such as mean-square error and peak signal-to-noise ratio, structural similarity index measure, and so on. This overview can help researchers to develop a better idea of what compression algorithms are used in certain domains and to find out if the presented performance parameters are of interest for the intended purpose.

A novel, rapid, and facile method for the colorimetric determination of calcium using micro-analytical paper-based devices (μ-PADs) was developed. The proposed analytical method utilizes the color differences developing, after the addition of calcium, on the surface of the devices because of the complexation reaction of calcium with Methylthymol Blue (MTB) at room temperature, in alkaline pH. The devices were manufactured with chromatographic paper, using wax barriers, and the analytical protocol was easily implemented without the need of any experimental apparatus except for a simple imaging device. The user must regulate the pH, add the solutions on the paper, and measure the color intensity of the formed Ca(II)-MTB complex with a flatbed scanner. The experimental conditions for optimum color development, the possible interfering substances, and the reliability of the paper devices in different preserving conditions were optimized, with satisfactory results. The method exhibited acceptable detection limits (2.9 mg L -1 ) with sufficiently good precision, which varied from 4.2% (intra-day) to 6.4% (inter-day). Saliva samples from healthy volunteers were successfully analyzed, and the calcium levels were calculated in the range of 30.71 to 84.15 mg L -1 .

Skin-based wearable devices have gained significant attention due to advancements in soft materials and thin-film technologies. Nevertheless, traditional wearable electronics often entail expensive and intricate manufacturing processes and rely on metal-based substrates that are susceptible to corrosion and lack flexibility. In response to these challenges, this paper has emerged with an alternative substrate for wearable electrodes due to its cost-effectiveness and scalability in manufacturing. Paper-based electrodes offer an attractive solution with their inherent properties of high breathability, flexibility, biocompatibility, and tunability. In this study, we introduce carbon nanotube-based paper composites (CPC) electrodes designed for the continuous detection of biopotential signals, such as electrooculography (EOG), electrocardiogram (ECG), and electroencephalogram (EEG). To prevent direct skin contact with carbon nanotubes, we apply various packaging materials, including polydimethylsiloxane (PDMS), Eco-flex, polyimide (PI), and polyurethane (PU). We conduct a comparative analysis of their signal-to-noise ratios in comparison to conventional gel electrodes. Our system demonstrates real-time biopotential monitoring for continuous health tracking, utilizing CPC in conjunction with a portable data acquisition system. The collected data are analyzed to provide accurate heart rates, respiratory rates, and heart rate variability metrics. Additionally, we explore the feasibility using CPC for sleep monitoring by collecting EEG signals.

A pipette-free and fully integrated device that can be used to accurately recognize the presence of infectious pathogens is an important and useful tool in point-of-care testing, particularly when aiming to decrease the unpredictable threats posed by disease outbreak. In this study, a paper device is developed to integrate the three main processes required for detecting infectious pathogens, including DNA extraction, loop-mediated isothermal amplification (LAMP), and detection. All key reagents, including sodium dodecyl sulfate (SDS), NaOH, LAMP reagents, and carmoisine, are placed on the paper device. The paper device is operated simply via sliding and folding without using any bulky equipment, and the results can be directly observed by the naked eye. The optimized concentrations of sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and carmoisine were found to be 0.1%, 0.1 M, and 0.5 mg/mL, respectively. The paper device was used to detect Enterococcus faecium at concentrations as low as 10 2 CFU/mL within 60 min. Also, E. faecium spiked in milk was successfully detected using the paper device, demonstrating the feasible application in real sample analysis.

In healthcare, new diagnostic tools that help in the diagnosis, prognosis, and monitoring of diseases rapidly and accurately are in high demand. For in-situ measurement of disease or infection biomarkers, point-of-care devices provide a dramatic speed advantage over conventional techniques, thus aiding clinicians in decision-making. During the last decade, paper-based analytical devices, combining paper substrates and electrochemical detection components, have emerged as important point-of-need diagnostic tools. This review highlights significant works on this topic over the last five years, from 2015 to 2019. The most relevant articles published in 2018 and 2019 are examined in detail, focusing on device fabrication techniques and materials applied to the production of paper fluidic and electrochemical cell architectures as well as on the final device assembly. Two main approaches were identified, that are, on one hand, those ones where the fabrication of the electrochemical cell is done on the paper substrate, where the fluidic structures are also defined, and, on the other hand, the fabrication of those ones where the electrochemical cell and liquid-driving paper component are defined on different substrates and then heterogeneously assembled. The main limitations of the current technologies are outlined and an outlook on the current technology status and future prospects is given., Competing Interests: The authors declare no conflict of interest.

In this paper, a low-cost resin-coated commercial-photo-paper substrate is used to design a printed reconfigurable multiband antenna. The two PIN diodes are used mainly to redistribute the surface current that provides reconfigurable properties to the proposed antenna. The antenna size of 40 mm × 40 mm × 0.44 mm with a partial ground, covers wireless and mobile bands ranging from 1.91 GHz to 6.75 GHz. The parametric analysis is performed to achieve optimized design parameters of the antenna. The U-shaped and C-shaped emitters are meant to function at 2.4 GHz and 5.9 GHz, respectively, while the primary emitter is designed to operate at 3.5 GHz. The proposed antenna achieved peak gain and radiation efficiency of 3.4 dBi and 90%, respectively. Simulated and measured results of the reflection coefficient, radiation pattern, gain, and efficiency show that the antenna design is in favorable agreement. Since the proposed antenna achieved wideband (1.91-6.75 GHz) using PIN diode configuration, using this technique the need for numerous electronic components to provide multiband frequency is avoided.

Norovirus is one of the leading causes of gastroenteritis, acute vomiting, intense diarrhoea, acute pain in the stomach, high fever, headaches, and body pain. Conventional methods of detection gave us very promising results but had disadvantages such as low sensitivity, cost ineffectiveness, reduced specificity and selectivity, etc. Therefore, biosensors can be a viable alternative device which can overcome all setbacks associated with the conventional method. An electrochemical sensor based on oxidized graphitic carbon nitride (Ox-g-C 3 N 4 ) modified electrochemical paper-based analytical device (ePAD) was fabricated for the detection of norovirus DNA. The synthesized Ox-g-C 3 N 4 nanosheets were characterized by field emission scanning electron microscopy (FESEM), X-ray Diffraction (XRD), UV-Vis spectroscopy and X-Ray Photoelectron Spectroscopy. The capture probe DNA (PDNA) modified electrodes were characterized by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). These two characterization techniques were also employed to find the optimal scan rate, response time and temperature of the fabricated sensor. The fabricated biosensor showed a limit of detection (LOD) of 100 fM. Furthermore, the specificity of the reported biosensor was affirmed by testing the response of capture probe DNA with oxidized graphitic carbon nitride (PDNA/Ox-g-C 3 N 4 ) modified ePAD on the introduction of a non-complimentary DNA. The fabricated ePAD sensor is easy to fabricate, cost effective and specific, and requires a minimum analysis time of 5 s.

Cancer is a major cause of mortality and morbidity worldwide. Detection and quantification of cancer biomarkers plays a critical role in cancer early diagnosis, screening, and treatment. Clinicians, particularly in developing countries, deal with high costs and limited resources for diagnostic systems. Using low-cost substrates to develop sensor devices could be very helpful. The interest in paper-based sensors with colorimetric detection increased exponentially in the last decade as they meet the criteria for point-of-care (PoC) devices. Cellulose and different nanomaterials have been used as substrate and colorimetric probes, respectively, for these types of devices in their different designs as spot tests, lateral-flow assays, dipsticks, and microfluidic paper-based devices (μPADs), offering low-cost and disposable devices. However, the main challenge with these devices is their low sensitivity and lack of efficiency in performing quantitative measurements. This review includes an overview of the use of paper for the development of sensing devices focusing on colorimetric detection and their application to cancer biomarkers. We highlight recent works reporting the use of paper in the development of colorimetric sensors for cancer biomarkers, such as proteins, nucleic acids, and others. Finally, we discuss the main advantages of these types of devices and highlight their major pitfalls.

Microfluidic paper-based analytical devices (µPADs) represent one of the promising green analytical strategies for low-cost and simple determination of various analytes. The actual task is the development of such devices for quantitation of antioxidants, e.g., flavonoids. In this paper, possibilities of a novel three-reagent µPAD including silver nitrate, 4-nitrophenyldiazonium tetrafluoroborate, and iron(III) chloride as reagents are assessed with respect to the determination of dihydroquercetin. It is shown that all the three reagents produce different colorimetric responses that can be detected by a mini-spectrophotometer-monitor calibrator or by a smartphone. The method is applicable to direct measuring high contents of dihydroquercetin (the linearity range is 0.026-1 mg mL -1 , and the limit of detection is 7.7 µg mL -1 ), which is favorable for many dietary supplements. The analysis of a food supplement was possible with the relative standard deviations of 9-26%, which is satisfactory for quantitative and semiquantitative determinations. It was found that plotting a calibration graph in 3D space of the three reagents' responses allows us to distinguish dihydroquercetin from its close structural analogue, quercetin.

Paper-based biosensors are a potential paradigm of sensitivity achieved via microporous spreading/microfluidics, simplicity, and affordability. In this paper, we develop decorated paper with graphene and conductive polymer (herein referred to as graphene conductive polymer paper-based sensor or GCPPS) for sensitive detection of biomolecules. Planetary mixing resulted in uniformly dispersed graphene and conductive polymer ink, which was applied to laser-cut Whatman filter paper substrates. Scanning electron microscopy and Raman spectroscopy showed strong attachment of conductive polymer-functionalized graphene to cellulose fibers. The GCPPS detected dopamine and cytokines, such as tumor necrosis factor-alpha (TNF-α), and interleukin 6 (IL-6) in the ranges of 12.5-400 µM, 0.005-50 ng/mL, and 2 pg/mL-2 µg/mL, respectively, using a minute sample volume of 2 µL. The electrodes showed lower detection limits (LODs) of 3.4 µM, 5.97 pg/mL, and 9.55 pg/mL for dopamine, TNF-α, and IL-6 respectively, which are promising for rapid and easy analysis for biomarkers detection. Additionally, these paper-based biosensors were highly selective (no serpin A1 detection with IL-6 antibody) and were able to detect IL-6 antigen in human serum with high sensitivity and hence, the portable, adaptable, point-of-care, quick, minute sample requirement offered by our fabricated biosensor is advantageous to healthcare applications.

Aptamers are short oligonucleotides designed to possess high binding affinity towards specific target compounds (ions, molecules, or cells). Due to their function and unique advantages, aptamers are considered viable alternatives to antibodies as biorecognition elements in bioassays and biosensors. On the other hand, paper-based devices (PADs) have emerged as a promising and powerful technology for the fabrication of low-cost analytical tools, mainly intended for on-site and point-of-care applications. The present work aims to provide a comprehensive overview of paper-based aptasensors. The review describes the fabrication methods and working principles of paper-based devices, the properties of aptamers as bioreceptors, the different modes of detection used in conjunction with aptasensing PADs, and representative applications for the detection of ions, small molecules, proteins, and cells. The future challenges and prospects of these devices are also discussed.

This paper presents the development of cheap and selective Paper-based Analytical Devices (PADs) for selective Pd(II) determination from very acidic aqueous solutions. The PADs were obtained by impregnating two cm-side squares of filter paper with an azoic ligand, (2-(tetrazolylazo)-1,8 dihydroxy naphthalene-3,6,-disulphonic acid), termed TazoC. The so-obtained orange TazoC-PADs interact quickly with Pd(II) in aqueous solutions by forming a complex purple-blue-colored already at pH lower than 2. The dye complexes no other metal ions at such an acidic media, making TazoC-PADs highly selective to Pd(II) detection. Besides, at higher pH values, other cations, for example, Cu(II) and Ni(II), can interact with TazoC through the formation of stable and pink-magenta-colored complexes; however, it is possible to quantify Pd(II) in the presence of other cations using a multivariate approach. To this end, UV-vis spectra of the TazoC-PADs after equilibration with the metal ions solutions were registered in the 300-800 nm wavelength range. By applying Partial Least Square regression (PLS), the whole UV-vis spectra of the TazoC-PADs were related to the Pd(II) concentrations both when present alone in solution and also in the presence of Cu(II) and Ni(II). Tailored PLS models obtained with matrix-matched standard solutions correctly predicted Pd(II) concentrations in unknown samples and tap water spiked with the metal cation, making the method promising for quick and economical sensing of Pd(II).

Adolescent idiopathic scoliosis (AIS) is a common disease that, in many cases, can be conservatively treated through bracing. High adherence to brace prescription is fundamental to gaining the maximum benefit from this treatment approach. Wearable sensors are available that objectively monitor the brace-wearing time, but their use, combined with other interventions, is poorly investigated. The aims of the current review are as follows: (i) to summarize the real compliance with bracing reported by studies using sensors; (ii) to find out the real brace wearing rate through objective electronic monitoring; (iii) to verify if interventions made to increase adherence to bracing can be effective according to the published literature. We conducted a systematic review of the literature published on Medline, EMBASE, CINAHL, Scopus, CENTRAL, and Web of Science. We identified 466 articles and included examples articles, which had a low to good methodological quality. We found that compliance a greatly varied between 21.8 and 93.9% (weighted average: 58.8%), real brace wearing time varied between 5.7 and 21 h per day (weighted average 13.3), and specific interventions seemed to improve both outcomes, with compliance increasing from 58.5 to 66% and brace wearing increasing from 11.9 to 15.1 h per day. Two comparative studies showed positive effects of stand-alone counseling and information on the sensors' presence when added to counseling. Sensors proved to be useful tools for objectively and continuously monitoring adherence to therapy in everyday clinical practice. Specific interventions, like the use of sensors, counseling, education, and exercises, could increase compliance. However, further studies using high-quality designs should be conducted in this field.

Home-based rehabilitation programs for older adults have demonstrated effectiveness, desirability, and reduced burden. However, the feasibility and effectiveness of balance-intervention training delivered through traditional paper-versus novel smartphone-based methods is unknown. Therefore, the purpose of this study was to evaluate if a home-based balance-intervention program could equally improve balance performance when delivered via smartphone or paper among adults over the age of 65. A total of 31 older adults were randomized into either a paper or phone group and completed a 4-week asynchronous self-guided balance intervention across 12 sessions for approximately 30 min per session. Baseline, 4-week, and 8-week walking and standing balance evaluations were performed, with exercise duration and adherence recorded. Additional self-reported measures were collected regarding the enjoyment, usability, difficulty, and length of the exercise program. Twenty-nine participants completed the balance program and three assessments, with no group differences found for any outcome measure. Older adults demonstrated an approximately 0.06 m/s faster gait velocity and modified balance strategies during walking and standing conditions following the intervention protocol. Participants further self-reported similar enjoyment, difficulty, and exercise effectiveness. Results of this study demonstrated the potential to safely deliver home-based interventions as well as the feasibility and effectiveness of delivering balance intervention through a smartphone-based application.

Paper-based sensors and assays have been highly attractive for numerous biological applications, including rapid diagnostics and assays for disease detection, food safety, and clinical care. In particular, the paper immunoassay has helped drive many applications in global health due to its low cost and simplicity of operation. This review is aimed at examining the fundamentals of the technology, as well as different implementations of paper-based assays and discuss novel strategies for improving their sensitivity, performance, or enabling new capabilities. These innovations can be categorized into using unique nanoparticle materials and structures for detection via different techniques, novel biological species for recognizing biomarkers, or innovative device design and/or architecture.

We present the development of an electrochemical paper-based analytical device (ePAD) for the detection of methamphetamine. Methamphetamine is a stimulant that young people use as an addictive narcotic, and it must be detected quickly since it may be hazardous. The suggested ePAD has the advantages of being simple, affordable, and recyclable. This ePAD was developed by immobilizing a methamphetamine-binding aptamer onto Ag-ZnO nanocomposite electrodes. The Ag-ZnO nanocomposites were synthesized via a chemical method and were further characterized via scanning electron microscopy, Fourier transform infrared spectroscopy, and UV-vis spectrometry in terms of their size, shape, and colloidal activity. The developed sensor showed a limit of detection of about 0.1 μg/mL, with an optimum response time of about 25 s, and its extensive linear range was between 0.01 and 6 μg/mL. The application of the sensor was recognized by spiking different beverages with methamphetamine. The developed sensor has a shelf life of about 30 days. This cost-effective and portable platform might prove to be highly successful in forensic diagnostic applications and will benefit those who cannot afford expensive medical tests.

The use of enzyme-based biosensors for the detection and quantification of analytes of interest such as contaminants of emerging concern, including over-the-counter medication, provides an attractive alternative compared to more established techniques. However, their direct application to real environmental matrices is still under investigation due to the various drawbacks in their implementation. Here, we report the development of bioelectrodes using laccase enzymes immobilized onto carbon paper electrodes modified with nanostructured molybdenum disulfide (MoS 2 ). The laccase enzymes were two isoforms (LacI and LacII) produced and purified from the fungus Pycnoporus sanguineus CS43 that is native to Mexico. A commercial purified enzyme from the fungus Trametes versicolor (TvL) was also evaluated to compare their performance. The developed bioelectrodes were used in the biosensing of acetaminophen, a drug widely used to relieve fever and pain, and of which there is recent concern about its effect on the environment after its final disposal. The use of MoS 2 as a transducer modifier was evaluated, and it was found that the best detection was achieved using a concentration of 1 mg/mL. Moreover, it was found that the laccase with the best biosensing efficiency was LacII, which achieved an LOD of 0.2 µM and a sensitivity of 0.108 µA/µM cm 2 in the buffer matrix. Moreover, the performance of the bioelectrodes in a composite groundwater sample from Northeast Mexico was analyzed, achieving an LOD of 0.5 µM and a sensitivity of 0.015 µA/µM cm 2 . The LOD values found are among the lowest reported for biosensors based on the use of oxidoreductase enzymes, while the sensitivity is the highest currently reported.

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.

The increase in anthropogenic pollution raises serious concerns regarding contamination of water bodies and aquatic species with potential implications on human health. Pharmaceutical compounds are a type of contaminants of emerging concern that are increasingly consumed and, thus, being frequently found in the aquatic environment. In this sense, an electrochemical sensor based on an unmodified and untreated carbon fiber paper (CPS-carbon paper sensor) was simply employed for the analysis of trimethoprim antibiotic in fish samples. First, the analytical conditions were thoroughly optimized in order for the CPS to achieve maximum performance in trimethoprim determination. Therefore, an electrolyte (0.1 M Britton-Robinson buffer) pH of 7 was selected and for square wave voltammetry parameters, optimum values of amplitude, frequency and step potential corresponded to 0.02 V, 50 Hz, and 0.015 V, respectively, whereas the deposition of analyte occurred at +0.7 V for 60 s. In these optimum conditions, the obtained liner range (0.05 to 2 µM), sensitivity (48.8 µA µM -1 cm -2 ), and LOD (0.065 µM) competes favorably with the commonly used GCE-based sensors or BDD electrodes that employ nanostructuration or are more expensive. The CPS was then applied for trimethoprim determination in fish samples after employing a solid phase extraction procedure based on QuEChERS salts, resulting in recoveries of 105.9 ± 1.8% by the standard addition method.

Traces of mercury ions in environmental water can harm humans and animals. Paper-based visual detection methods have been widely developed for the rapid detection of mercury ions; however, existing methods are not sensitive enough to be used in real environments. Here, we developed a novel, simple and effective visual fluorescent sensing paper-based chip for the ultrasensitive detection of mercury ions in environmental water. CdTe-quantum-dots-modified silica nanospheres were firmly absorbed by and anchored to the fiber interspaces on the paper's surface to effectively avoid the unevenness caused by liquid evaporation. The fluorescence of quantum dots emitted at 525 nm can be selectively and efficiently quenched with mercury ions, and the ultrasensitive visual fluorescence sensing results attained using this principle can be captured using a smartphone camera. This method has a detection limit of 2.83 µg/L and a fast response time (90 s). We successfully achieved the trace spiking detection of seawater (from three regions), lake water, river water and tap water with recoveries in the range of 96.8-105.4% using this method. This method is effective, low-cost, user-friendly and has good prospects for commercial application. Additionally, the work is expected to be utilized in the automated big data collection of large numbers of environmental samples.

In this paper, the paper cups were used as the research objects, and the machine vision detection technology was combined with different image processing techniques to investigate a non-contact optical automatic detection system to identify the defects of the manufactured paper cups. The combined ring light was used as the light source, an infrared (IR) LED matrix panel was used to provide the IR light to constantly highlight the outer edges of the detected objects, and a multi-grid pixel array was used as the image sensor. The image processing techniques, including the Gaussian filter, Sobel operator, Binarization process, and connected component, were used to enhance the inspection and recognition of the defects existing in the produced paper cups. There were three different detection processes for paper cups, which were divided into internal, external, and bottom image acquisition processes. The present study demonstrated that all the detection processes could clearly detect the surface defect features of the manufactured paper cups, such as dirt, burrs, holes, and uneven thickness. Our study also revealed that the average time for the investigated Automatic Optical Detection to detect the defects on the paper cups was only 0.3 s.

For the first time, a novel NiFe 2 O 4 /paper-based magnetoelastic (ME) biosensor was developed for rapid, sensitive, and portable detection of human serum albumin (HSA). Due to the uniquely magnetoelastic effect of NiFe 2 O 4 nanoparticles and the excellent mechanical properties of the paper, the paper-based ME biosensor transforms the surface stress signal induced by the specific binding of HSA and antibody modified on the paper into the electromagnetic signal. The accumulated binding complex generates a compressive stress on the biosensor surface, resulting in a decrease in the biosensor's static magnetic permeability, which correlates to the HSA concentrations. To improve the sensitivity of the biosensor, the concentration of NiFe 2 O 4 nanofluid and the impregnated numbers of the NiFe 2 O 4 nanofluid-impregnated papers were optimized. The experimental results demonstrated that the biosensor exhibited a linear response to HSA concentrations ranging from 10 μg∙mL -1 to 200 μg∙mL -1 , with a detection limit of 0.43 μg∙mL -1 , which is significantly lower than the minimal diagnosis limit of microalbuminuria. The NiFe 2 O 4 /paper-based ME biosensor is easy to fabricate, and allows the rapid, highly-sensitive, and selective detection of HSA, providing a valuable analytical device for early monitoring and clinical diagnosis of microalbuminuria and nephropathy. This study shows the successful integration of the paper-based biosensor and the ME sensing analytical method will be a highly-sensitive, easy-to-use, disposable, and portable alternative for point-of-care monitoring.

Although the established ELISA-based sensing platforms have many benefits, the importance of cytokine and cancer biomarkers detection for point-of-care diagnostics has propelled the search for more specific, sensitive, simple, accessible, yet economical sensor. Paper-based biosensor holds promise for future in-situ applications and can provide rapid analysis and data without the need to conduct in a laboratory. Electrochemical detection plays a vital role in interpreting results obtained from qualitative assessment to quantitative determination. In this review, various factors affecting the design of an electrochemical paper-based biosensor are highlighted and discussed in depth. Different detection methods, along with the latest development in utilizing them in cytokine and cancer biomarkers detection, are reviewed. Lastly, the fabrication of portable electrochemical paper-based biosensor is ideal in deliberating positive societal implications in developing countries with limited resources and accessibility to healthcare services.

Since the determination of the high toxicity of bisphenol A, alternative structures for bisphenols have been synthesized, resulting in bisphenols C, E, F, S, and Z. These bisphenols have replaced bisphenol A in plastic bottles, toys, and cans used for preserving food. Later, the toxicity and negative effects of all of these bisphenols on people's health were proven. Therefore, there is a need for a fast ultrasensitive screening method that is able to detect the presence of these bisphenols in any condition directly from food samples. This paper presented a disposable device based on the utilization of a 2D disposable paper stochastic sensor for the fast ultrasensitive screening of food samples for bisphenols A, C, E, F, S, and Z. The 2D disposable sensor was obtained by the deposition of graphene and silver nanolayers on paper using cold plasma. Furthermore, the active side of the sensor was modified using 2,3,7,8,12,13,17,18-octaethyl-21H,23H Mn porphyrin. The limits of quantification of these bisphenols were 1 fmol L -1 for bisphenols C and E, 10 fmol L -1 for bisphenols A and F, 10 pmol L -1 for bisphenol S, and 1 pmol L -1 for bisphenol Z. The recoveries of these bisphenols in milk, canned fruits, vegetables, and fish were higher than 99.00% with RSD (%) values lower than 1.50%.

Soil tests for plant-available phosphorus (P) are suggested to provide offsite P analysis required to monitor P fertilizer application and reduce P losses to downstream water. However, procedural and cost limitations of current soil phosphate tests have restricted their widespread use and have made them accessible only in laboratories. This study proposes a novel paper-based reagentless electrochemical soil phosphate sensor to extract and detect soil phosphate using an inexpensive and simple approach. In this test, concentrated Mehlich-3 and molybdate ions were impregnated in filter paper, which served as the phosphate extraction and reaction zone, and was followed by electrochemical detection using cyclic voltammetry signals. Soil samples from 22 sampling sites were used to validate this method against inductively coupled plasma optical emission spectroscopy (ICP) soil phosphate tests. Regression and correlation analyses showed a significant relationship between phosphate determinations by ICP and the proposed method, delivering a correlation coefficient, r, of 0.98 and a correlation slope of 1.02. The proposed approach provided a fast, portable, low-cost, accessible, reliable, and single-step test to extract and detect phosphate simultaneously with minimum waste (0.5 mL per sample), which made phosphate characterization possible in the field.

The role of humidity sensors in different industries and field applications, such as agriculture, food monitoring, biomedical equipment, heating, and ventilation, is well known. However, most commercially available humidity sensors are based on polymers or electronic materials that are not degradable and thus contribute to electronic waste. Here, we report a low-cost, flexible, easy-to-fabricate, and eco-friendly parallel-plate capacitive humidity sensor for field applications. The sensor is fabricated from copper tape and tissue paper, where copper tape is used to create the plates of the capacitor, and tissue paper is used as a dielectric sensing layer. Along with the low cost, the high sensitivity, better response and recovery times, stability, and repeatability make this sensor unique. The sensor was tested for relative humidity (RH), ranging from 40% to 99%, and the capacitance varied linearly with RH from 240 pF to 720 pF, as measured by an Arduino. The response time of the sensor is ~1.5 s, and the recovery time is ~2.2 s. The experiment was performed 4-5 times on the same sensor, and repeatable results were achieved with an accuracy of ±0.1%. Furthermore, the sensor exhibits a stable response when tested at different temperatures. Due to the above advantages, the presented sensor can find ready applications in different areas.

The distinct properties and affordances of paper provide benefits that enabled paper to maintain an important role in the digital age. This is so much so, that some pen-paper interaction has been imitated in the digital world with touchscreens and stylus pens. Because digital medium also provides several advantages not available to physical paper, there is a clear benefit to merge the two mediums. Despite the plethora of concepts, prototypes and systems to digitise handwritten information on paper, these systems require specially prepared paper, complex setups and software, which can be used solely in combination with paper, and, most importantly, do not support the concurrent precise interaction with both mediums (paper and touchscreen) using one pen only. In this paper, we present the design, fabrication and evaluation of the Hybrid Stylus. The Hybrid Stylus is assembled with the infinity pencil tip (nib) made of graphite and a specially designed shielded tip holder that is attached to an active stylus. The stylus can be used for writing on a physical paper, while it still maintains all the features needed for tablet interaction. Moreover, the stylus also allows simultaneous digitisation of handwritten information on the paper when the paper is placed on the tablet screen. In order to evaluate the concept, we also add a user-friendly manual alignment of paper position on the underlying tablet computer The evaluation demonstrates that the system achieves almost perfect digitisation of strokes (98.6% of strokes were correctly registered with only 1.2% of ghost strokes) whilst maintaining excellent user experience of writing with a pencil on the paper.

The direct integration of paper-based microfluidic fuel cells (μFC's) toward creating autonomous lateral flow assays has attracted attention. Here, we show that an air-breathing paper-based μFC could be used as a power supply in pregnancy tests by oxidizing the human urine used for the diagnosis. We present an air-breathing paper-based μFC connected to a pregnancy test, and for the first time, as far as we know, it is powered by human urine without needing any external electrolyte. It uses TiO2-Ni as anode and Pt/C as cathode; the performance shows a maximum value of voltage and current and power densities of ∼0.96 V, 1.00 mA cm-2, and 0.23 mW cm-2, respectively. Furthermore, we present a simple design of a paper-based μFC's stack powered with urine that shows a maximum voltage and maximum current and power densities of ∼1.89 V, 2.77 mA cm-2 and 1.38 mW cm-2, respectively, which powers the display of a pregnancy test allowing to see the analysis results.

The general objective of Analytical Chemistry, nowadays, is to obtain best-quality information in the shortest time to contribute to the resolution of real problems. In this regard, electrochemical biosensors are interesting alternatives to conventional methods thanks to their great characteristics, both those intrinsically analytical (precision, sensitivity, selectivity, etc.) and those more related to productivity (simplicity, low costs, and fast response, among others). For many years, the scientific community has made continuous progress in improving glucose biosensors, being this analyte the most important in the biosensor market, due to the large amount of people who suffer from diabetes mellitus . The sensitivity of the electrochemical techniques combined with the selectivity of the enzymatic methodologies have positioned electrochemical enzymatic sensors as the first option. This review, focusing on the electrochemical determination of glucose using paper-based analytical devices, shows recent approaches in the use of paper as a substrate for low-cost biosensing. General considerations on the principles of enzymatic detection and the design of paper-based analytical devices are given. Finally, the use of paper in enzymatic electrochemical biosensors for glucose detection, including analytical characteristics of the methodologies reported in relevant articles over the last years, is also covered.

Hydrogen sulfide (H 2 S) is known to participate in bacteria-induced inflammatory response in periodontal diseases. Therefore, it is necessary to quantify H 2 S produced by oral bacteria for diagnosis and treatment of oral diseases including halitosis and periodontal disease. In this study, we introduce a paper-based colorimetric assay for detecting bacterial H 2 S utilizing silver/Nafion/polyvinylpyrrolidone membrane and a 96-well microplate. This H 2 S-sensing paper showed a good sensitivity (8.27 blue channel intensity/μM H 2 S, R 2 = 0.9996), which was higher than that of lead acetate paper (6.05 blue channel intensity/μM H 2 S, R 2 = 0.9959). We analyzed the difference in H 2 S concentration released from four kinds of oral bacteria ( Eikenella corrodens , Streptococcus sobrinus , Streptococcus mutans , and Lactobacillus casei ). Finally, the H 2 S level in Eikenella corrodens while varying the concentration of cysteine and treatment time was quantified. This paper-based colorimetric assay can be utilized as a simple and effective tool for in vitro screening of H 2 S-producing ability of many bacteria as well as salivary H 2 S analysis.

Since the beginning of the COVID-19 pandemic, many works have been published proposing solutions to the problems that arose in this scenario. In this vein, one of the topics that attracted the most attention is the development of computer-based strategies to detect COVID-19 from thoracic medical imaging, such as chest X-ray (CXR) and computerized tomography scan (CT scan). By searching for works already published on this theme, we can easily find thousands of them. This is partly explained by the fact that the most severe worldwide pandemic emerged amid the technological advances recently achieved, and also considering the technical facilities to deal with the large amount of data produced in this context. Even though several of these works describe important advances, we cannot overlook the fact that others only use well-known methods and techniques without a more relevant and critical contribution. Hence, differentiating the works with the most relevant contributions is not a trivial task. The number of citations obtained by a paper is probably the most straightforward and intuitive way to verify its impact on the research community. Aiming to help researchers in this scenario, we present a review of the top-100 most cited papers in this field of investigation according to the Google Scholar search engine. We evaluate the distribution of the top-100 papers taking into account some important aspects, such as the type of medical imaging explored, learning settings, segmentation strategy, explainable artificial intelligence (XAI), and finally, the dataset and code availability.

The electrocardiogram (ECG) signal describes the heart's electrical activity, allowing it to detect several health conditions, including cardiac system abnormalities and dysfunctions. Nowadays, most patient medical records are still paper-based, especially those made in past decades. The importance of collecting digitized ECGs is twofold: firstly, all medical applications can be easily implemented with an engineering approach if the ECGs are treated as signals; secondly, paper ECGs can deteriorate over time, therefore a correct evaluation of the patient's clinical evolution is not always guaranteed. The goal of this paper is the realization of an automatic conversion algorithm from paper-based ECGs (images) to digital ECG signals. The algorithm involves a digitization process tested on an image set of 16 subjects, also with pathologies. The quantitative analysis of the digitization method is carried out by evaluating the repeatability and reproducibility of the algorithm. The digitization accuracy is evaluated both on the entire signal and on six ECG time parameters (R-R peak distance, QRS complex duration, QT interval, PQ interval, P-wave duration, and heart rate). Results demonstrate the algorithm efficiency has an average Pearson correlation coefficient of 0.94 and measurement errors of the ECG time parameters are always less than 1 mm. Due to the promising experimental results, the algorithm could be embedded into a graphical interface, becoming a measurement and collection tool for cardiologists.

Microneedle (MN) is a novel technique of the biomedical engineering field because of its ability to evaluate bioinformation via minimal invasion. One of the urgent requirements for ground-breaking health care monitoring is persistent monitoring. Hollow microneedles are extremely attractive to extract skin interstitial fluid (ISF) for analysis, which makes them perfect for sensing biomarkers and facilitating diagnosis. Nevertheless, its intricate fabrication process has hampered its extensive application. The present research demonstrates an easy one-step preparation approach for hollow MNs on the foundation of the refraction index variations of polyethylene glycol diacrylate (PEGDA) in the process of photopolymerization. The fabricated hollow microneedle exhibited ideal mechanical characteristics to penetrate the skin. Hydrodynamic simulations showed that the liquid was risen in a hollow microneedle by capillary force. Furthermore, a paper-based glucose sensor was integrated with the hollow microneedle. We also observed that the MN array smoothly extracted ISF in vitro and in vivo by capillary action. The outcomes displayed the applicability of the MN patch to persistent blood glucose (GLU) monitoring, diagnosis-related tests for patients and pre-diabetic individuals.

The paper describes the development of an electrochemical sensor to be used for the determination of synthetic food colorants such as Sunset Yellow FCF (SY) and Tartrazine (TZ). The sensor is a carbon paper (CP) electrode, manufactured by using hot lamination technology and volume modified with fine-grained graphite powder (GrP). The sensor (GrP/CP) was characterized by scanning electron microscopy, energy dispersive spectrometry, electrochemical impedance analysis, cyclic, linear sweep and differential pulse voltammetry. The mechanism of SY and TZ electrochemical oxidation on GrP/CP was studied. The developed sensor has good electron transfer characteristics and low electron resistance, high sensitivity and selectivity. Applying the differential pulse mode, linear dynamic ranges of 0.005-1.0 μM and 0.02-7.5 μM with limits of detection of 0.78 nM and 8.2 nM for SY and TZ, respectively, were obtained. The sensor was used to detect SY and TZ in non-alcoholic and alcoholic drinks. The results obtained from drink analysis prove good reproducibility (RSD ≤ 0.072) and accuracy (recovery 96-104%).

To stabilize the detection signal of palladium-based hydrogen sensors on paper substrates, a graphite intermediate layer was painted on the surface of paper. The graphite-on-paper (GOP) substrate offers advantages such as good thermo-electrical conductivity, low cost, and uncomplicated preparation technology. Quasi-1-dimensional palladium (Pd) thin films with 8 nm and 60 nm thicknesses were deposited on the GOP substrates using the vacuum evaporation technique. Thanks to the unique properties of the GOP substrate, a continuous Pd microfiber network structure appeared after deposition of the ultra-thin Pd film. Additionally, the sensing performance of the palladium-based hydrogen sensor was not affected, whether using GOP or paper substrate at 25 °C. Surprisingly, heating-induced loss of sensitivity was restrained due to the increased electrical conductivity of the GOP substrate at 50 °C.

With a growing number of electronic devices surrounding our daily life, it becomes increasingly important to create solutions for clear and simple communication and interaction at the human machine interface (HMI). Haptic feedback solutions play an important role as they give a clear direct link and response to the user. This work demonstrates multifunctional haptic feedback devices based on fully printed piezoelectric transducers realized with functional polymers on thin paper substrate. The devices are flexible; lightweight and show very high out-of-plane deflection of 213 µm at a moderate driving voltage of 50 V rms (root mean square) achieved by an innovative multilayer design with up to five individually controllable active layers. The device creates a very clear haptic sensation to the human skin with a blocking force of 0.6 N at the resonance frequency of 320 Hz, which is located in the most sensitive range of the human fingertip. Additionally the transducer generates audible information above two kilohertz with a remarkable high sound pressure level. Thus the paper-based approach can be used for interactive displays in combination with touch sensation; sound and color prints. The work gives insights into the manufacturing process; the electrical characteristics; and an in-depth analysis of the 3D deflection of the device under variable conditions.

In recent years, the wide application of deep neural network models has brought serious risks of intellectual property rights infringement. Embedding a watermark in a network model is an effective solution to protect intellectual property rights. Although researchers have proposed schemes to add watermarks to models, they cannot prevent attackers from adding and overwriting original information, and embedding rates cannot be quantified. Therefore, aiming at these problems, this paper designs a high embedding rate and tamper-proof watermarking scheme. We employ wet paper coding (WPC), in which important parameters are regarded as wet blocks and the remaining unimportant parameters are regarded as dry blocks in the model. To obtain the important parameters more easily, we propose an optimized probabilistic selection strategy (OPSS). OPSS defines the unimportant-level function and sets the importance threshold to select the important parameter positions and to ensure that the original function is not affected after the model parameters are changed. We regard important parameters as an unmodifiable part, and only modify the part that includes the unimportant parameters. We selected the MNIST, CIFAR-10, and ImageNet datasets to test the performance of the model after adding a watermark and to analyze the fidelity, robustness, embedding rate, and comparison schemes of the model. Our experiment shows that the proposed scheme has high fidelity and strong robustness along with a high embedding rate and the ability to prevent malicious tampering.

This study aimed to design and fabricate planar constricted-expanded structures that are integrated into paper-based channels in order to enhance their chaotic advection and improve their mixing performance. Chromatography papers were used to print paper-based microfluidics using a solid-wax printer. Three different constricted-expanded structures-i.e., zigzag, crossed, and curved channels-were designed in order to evaluate their mixing performance in comparison with that of straight channels. A numerical simulation was performed in order to investigate the mixing mechanism, and to examine the ways in which the planar constricted-expanded structures affected the flow patterns. The experimental and numerical results indicated that the proposed structures can successfully mix confluents. The experimental results revealed that the mixing indices ( σ ) rose from the initial 20.1% (unmixed) to 34.5%, 84.3%, 87.3%, and 92.4% for the straight, zigzag, curved, and cross-shaped channels, respectively. In addition, the numerical calculations showed a reasonable agreement with the experimental results, with a variation in the range of 1.0-11.0%. In future, we hope that the proposed passive paper-based mixers will be a crucial component in the application of paper-based microfluidic devices.