Microfluidic Paper-based Analytical Devices Research Articles

Microfluidic paper-based analytical device integrated with Fe@ZnS:MIP for colorimetric detection of antibiotics

Microfluidic paper-based analytical device integrated with Fe@ZnS:MIP for colorimetric detection of antibiotics

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

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

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

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

Disposable Device for Bacterial Vaginosis Detection.

Due to the increasing demand for clinical testing of infectious diseases at the point-of-care, the global market claims alternatives for rapid diagnosis tools such as disposable biosensors, avoiding the need for specialized laboratories and skilled personnel. Bacterial vaginosis (BV) is an infectious disease that commonly affects reproductive-age women and predisposes the infection of sexually transmitted diseases. Especially in asymptomatic cases, BV can lead to pelvic inflammatory conditions, postpartum endometritis, and preterm labor. Conventionally, BV diagnosis involves the microscopic analysis of vaginal swab samples; it thus requires highly trained personnel. In response, we report a novel microfluidic paper-based analytical device for BV diagnosis. Sialidase, a biomarker overexpressed in BV, was detected by exploiting an immunosensing mechanism previously discovered by our team. This technology employs a graphene oxide-coated surface as a quencher of fluorescence; the fluorescence of the immunoprobes that do not experiment immunoreactions (antibody-antigen) are deactivated by graphene oxide via non-radiative energy transfer, whereas those immunoprobes undergoing immunoreactions preserve their photoluminescence due to the distance and the low affinity between the immunocomplex and the graphene oxide-coated surface. Our paper-based test was typically carried out within 20 min, and the sample volume was 6 μL. Besides, it was tested with 14 vaginal swabs specimens to discriminate clinical samples of women with normal microbiota from those with BV. Our disposable device represents a new tool to prevent the consequences of BV.

Paper-Based Laser-Pyrolyzed Electrofluidics: An Electrochemical Platform for Capillary-Driven Diagnostic Bioassays.

Microfluidic paper-based analytical devices (µPADs) are indispensable tools for disease diagnostics. The integration of electronic components into µPADs enables new device functionalities and facilitates the development of complex quantitative assays. Unfortunately, current electrode fabrication methods often hinder capillary flow, considerably restricting µPAD design architectures. Here, laser-induced graphenization is presented as an approach to fabricate porous electrodes embedded into cellulose paper. The resulting electrodes not only have high conductivity and electrochemical activity, but also retain wetting properties for capillary transport. Paper-based electrofluidics, including a lateral flow device for injection analysis of alkaline phosphatase in serum and a vertical flow device for quantitative detection of HPV16 with a CRISPR-based assay are demonstrated. It is expected that this platform will streamline the development of diagnostic devices that combine the operational simplicity of colorimetric lateral flow tests with the added benefits and possibilities offered by electronic signaling.

Graphene oxide modified cellulose paper-based device: A novel platform for cooking oil quality evaluation

A novel platform of microfluidic paper-based analytical device (μPAD) with colorimetric acid-base titration for cooking oil quality evaluation was created and oleic acid was selected as a model analyte. Graphene oxide (GO) was used to modify cellulose paper for improved visualization and increased hydrophobicity of the colorimetric sensor. μPAD with colorimetric acid-base titration was prepared by immobilizing phenolphthalein and primary standard of Na2CO3 on GO-modified cellulose paper and applied for oleic acid detection. The neutralization of oleic acid on the cellulose paper was illustrated by color disappearance from pink to colorless. This platform provided a linearity of (0–2% v/v), covering a cut-off oleic acid level (1.0%) for evaluation of cooking oil quality. This device, designed with 3D-μPAD, created the automatic lateral-flow channel for semi-quantitative analysis of cooking oil without instrumental requirement. The content of oleic acid equated to the number of pink-detection area disappearing. Ultimately, this device was successfully applied for cooking oil quality evaluation in real waste cooking oil samples.

Microfluidic paper-based analytical devices for simultaneous detection of oxidative potential and copper in aerosol samples.

The potential reach of point-of-care (POC) diagnostics into daily routines for exposure to reactive oxygen species (ROS) and Cu in aerosolized particulate matter (PM) demands that microfluidic paper-based analytical devices (μPADs) take into consideration the simple detection of these toxic PM components. Here, we propose μPADs with a dual-detection system for simultaneous ROS and Cu(II) detection. For colorimetric ROS detection, the glutathione (GSH) assay with a folding design to delay the reaction yielded complete ROS and GSH oxidation, and improved homogeneity of color development relative to using the lateral flow pattern. For electrochemical Cu(II) determination, 1,10-phenanthroline/Nafion modified graphene screen-printed electrodes showed ability to detect Cu(II) down to pg level being low enough to be applied to PM analysis. No intra- and inter-interference affecting both systems were found. The proposed μPADs obtained LODs for 1,4-naphthoquinone (1,4-NQ), used as the ROS representative, and Cu(II) of 8.3 ng and 3.6 pg, respectively and linear working ranges of 20 to 500 ng for ROS and 1 × 10-2 to 2 × 102 ng for Cu(II). Recovery of the method was between 81.4 and 108.3% for ROS and 80.5-105.3% for Cu(II). Finally, the sensors were utilized for simultaneous ROS and Cu(II) determination in PM samples and the results statistically agreed with those using the conventional methods at 95% confidence.

Modeling of Paper-Based Bi-Material Cantilever Actuator for Microfluidic Biosensors.

This research explores the dynamics of a fluidically loaded Bi-Material cantilever (B-MaC), a critical component of μPADs (microfluidic paper-based analytical devices) used in point-of-care diagnostics. Constructed from Scotch Tape and Whatman Grade 41 filter paper strips, the B-MaC's behavior under fluid imbibition is examined. A capillary fluid flow model is formulated for the B-MaC, adhering to the Lucas-Washburn (LW) equation, and supported by empirical data. This paper further investigates the stress-strain relationship to estimate the modulus of the B-MaC at various saturation levels and to predict the behavior of the fluidically loaded cantilever. The study shows that the Young's modulus of Whatman Grade 41 filter paper drastically decreases to approximately 20 MPa (about 7% of its dry-state value) upon full saturation. This significant decrease in flexural rigidity, in conjunction with the hygroexpansive strain and coefficient of hygroexpansion (empirically deduced to be 0.008), is essential in determining the B-MaC's deflection. The proposed moderate deflection formulation effectively predicts the B-MaC's behavior under fluidic loading, emphasizing the measurement of maximum (tip) deflection using interfacial boundary conditions for the B-MaC's wet and dry regions. This knowledge of tip deflection will prove instrumental in optimizing the design parameters of B-MaCs.

Use of scanners for colorimetric analysis of microfluidic paper-based analytical devices (µPADs): A practical guide

Office style scanners are often used by researchers for the colorimetric detection in the development of microfluidic paper-based analytical devices (µPADs). However, little attention has been paid to nuances in calibration curve responses caused by varying settings of a single image-capturing device. Hence, in this study, the effect of parameters, such as the scanner operation conditions, the method of data analysis used (e.g., choice of colour space system, suitable channel), and the influence of the physical characteristics of the µPAD (e.g., template colour, type of lamination pouches used) on the colorimetric detection were evaluated for the first time.Combinations of the scanning parameters were thoroughly examined and their impact on the quality of a mock calibration curve assessed. The most sensitive calibration curve was achieved only by the selection of the optimal scanning conditions combined with an appropriate choice of colorimetric system. A flatbed scanner, used in this study, showed greater variation of signal with varying scanning setup, whereas a portable scanner provided nearly identical responses under diverse scanning conditions. Gamma correction, scanning resolution, region of interest (ROI) surrounding colour (i.e., template colour), bit-depth, image file type, and the number of analysed pixels were found not to markedly affect the dynamics of the calibration curve when colour in the ROI is homogeneous. Parameters that had an effect on the colorimetric detection were the scanner settings, colour of the blank, image acquisition mode (colour vs greyscale), and µPAD covering (e.g., lamination). Based on these findings, generic guidelines for optimal data acquisition and µPAD manipulation with respect to µPAD calibration have been formulated.

Cover Feature: Enhanced Chemiluminescence of a Superior Luminol Derivative Provides Sensitive Smartphone‐Based Point‐of‐Care Testing with Enzymatic μPAD (Anal. Sens. 4/2023)

The cover image illustrates the principle of a newly developed chemiluminescence (CL) microfluidic paper-based analytical device (μPAD) that enables highly sensitive detection of any catalytic process in which H2O2 is produced. Through the use of a new, sensitive CL luminophore cell phone camera detection allows quantification on-site with the same sensitivity as afforded through a CCD camera in the lab. One-step reactions to time the catalytic reactions can easily be realized through a moveable barrier if desired. Lactate is detected with μM detection limits and three orders of magnitude dynamic range in sweat, the luminophore reaches a pM detection limit. Cover design by Simone Rink. More information can be found in the Research Article by Antje Baeumner and co-workers .

A novel photonic chemosensor for rapidly detecting synthetic dyes in orange juice using colorimetric and spectrophotometric methods

Synthetic dyes must be monitored and regulated. We aimed to develop a novel photonic chemosensor for rapidly monitoring synthetic dyes based on colorimetric (chemical interactions with optical probes using microfluidic paper-based analytical devices) and UV–Vis spectrophotometric methods. Various types of gold and silver nanoparticles were surveyed to identify the targets. In the presence of silver nanoprisms, the naked eye could visualize the unique and distinctive color changes of Tartrazine (Tar) to green and Sunset Yellow (Sun) to brown; UV–Vis spectrophotometry validated the results. The developed chemosensor showed linear ranges of 0.07–0.3 mM and 0.05–0.2 mM for Tar and Sun, respectively. Sources of interference had minimal effects, confirming the appropriate selectivity of the developed chemosensor. Our novel chemosensor demonstrated excellent analytical performance for measuring Tar and Sun in several types of orange juice as real samples, confirming its incredible potential for use in the food industry.

Mickey mouse-shaped laminated paper-based analytical device in simultaneous total cholesterol and glucose determination in whole blood

The microfluidic paper-based analytical device (μPAD) platform is gaining attention as a low-cost, portable, and disposable detection tool. However, the limitations of traditional fabrication methods include poor reproducibility and the use of hydrophobic reagents. In this study, an in-house computer-controlled X–Y knife plotter and pen plotter were used to fabricate μPADs, resulting in a simple, more rapid, reproducible process that consumes less volume of reagents. The μPADs were laminated to increase mechanical strength and reduce sample evaporation during analysis. The resulting laminated paper-based analytical device (LPAD) was used to simultaneously determine glucose and total cholesterol in whole blood using the LF1 membrane as a sample zone. The LF1 membrane selectively separates plasma from whole blood by size exclusion and yields plasma for further enzymatic reaction steps while retaining blood cells and larger proteins. The i1 Pro 3 mini spectrophotometer directly detected color on the LPAD. The results were clinically relevant and in agreement with hospital methods, with a detection limit of 0.16 mmol L⁻1 for glucose and 0.57 mmol L⁻1 for TC. The LPAD retained color intensity after 60 days of storage. The LPAD offers a low-cost, high-performance option for chemical sensing devices and expands the applicability of markers for diagnosing whole blood samples.

Semi-quantitative microfluidic paper-based analytical device for ionic silica detection.

Silicate ions (SiO32-), or ionic silica, are known to cause silica scaling in industrial water applications when excess amounts are present; hence, concentrations must be monitored and kept at a constant low level. Ionic silica is conventionally measured by spectrophotometry in the form of its silicomolybdic complex based on the molybdenum blue reaction, but the operation process is complicated and not suitable for on-site detection. To solve these issues, microfluidic paper-based analytical devices (µPADs) have been gaining attention as portable, low-cost analytical devices suitable for on-site detection. The foldable origami type device described in this work enabled silica detection based on the molybdenum blue reaction, in the range of 50-1000mg/L, with a practically detectable lowest concentration of 50mg/L. The device showed selectivity for silicate ions and stability over 21days when stored at 4°C. The semi-quantitative analytical performance makes the proposed paper-based device attractive for on-site industrial monitoring.

Iodometric Titration on Microfluidic Paper-Based Analytical Devices for Determination of Ascorbic Acid: A Laboratory Experiment for Chemical Education Undergraduates

Iodometric Titration on Microfluidic Paper-Based Analytical Devices for Determination of Ascorbic Acid: A Laboratory Experiment for Chemical Education Undergraduates

Nanomaterials integrated with microfluidic paper-based analytical devices for enzyme-free glucose quantification

In this study, nanomaterials capable of enzyme-free glucose quantification and colorimetric readout are integrated into a microfluidic paper-based analytical devices (μPADs). Gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) were utilized as a peroxidase-like nanozyme and a colorimetric probe to achieve glucose monitoring. In this developed device, glucose is oxidized by AuNPs to generate hydrogen peroxide (H2O2), which flows in the paper microchannels toward detection zones. H2O2 then etches the immobilized AgNPs to induce a color change. The intensity of color change is easily monitored using a smartphone application. Following method optimization, we obtained a linear range from 0.50 to 10.0 mmol L−1 (R2 = 0.9921) and a detection limit (LOD) of 340.0 μmol L−1. This falls in the clinically relevant range for glucose monitoring and diabetes diagnosis in humans. In addition, the total analysis time is just 20 min, which is significantly less than the same experiment performed in the solution phase. Also, our method is markedly selective; other substrates do not interfere. The recovery test in human control samples was in the range of 98.47–102.34% and the highest relative standard deviation (RSD) was 3.58%. The enzyme-free approach for glucose sensing is highly desirable for diabetes diagnosis as it replaces the more expensive enzyme with cheaper nanomaterials. Furthermore, since nanomaterials are more environmentally stable compared to enzymes, it has the potential for widespread deployment as point-of-care diagnostics (POC) in resource-limited settings.

Deep learning-assisted ultra-accurate smartphone testing of paper-based colorimetric ELISA assays

Smartphone has long been considered as one excellent platform for disease screening and diagnosis, especially when combined with microfluidic paper-based analytical devices (μPADs) that feature low cost, ease of use, and pump-free operations. In this paper, we report a deep learning-assisted smartphone platform for ultra-accurate testing of paper-based microfluidic colorimetric enzyme-linked immunosorbent assay (c-ELISA). Different from existing smartphone-based μPAD platforms, whose sensing reliability is suffered from uncontrolled ambient lighting conditions, our platform is able to eliminate those random lighting influences for enhanced sensing accuracy. We first constructed a dataset that contains c-ELISA results (n=2048) of rabbit IgG as the model target on μPADs under eight controlled lighting conditions. Those images are then used to train four different mainstream deep learning algorithms. By training with these images, the deep learning algorithms can well eliminate the influences of lighting conditions. Among them, the GoogLeNet algorithm gives the highest accuracy (>97%) in quantitative rabbit IgG concentration classification/prediction, which also provides 4% higher area under curve (AUC) value than that of the traditional curve fitting results analysis method. In addition, we fully automate the whole sensing process and achieve the "image in, answer out" to maximize the convenience of the smartphone. A simple and user-friendly smartphone application has been developed that controls the whole process. This newly developed platform further enhances the sensing performance of μPADs for use by laypersons in low-resource areas and can be facilely adapted to the real disease protein biomarkers detection by c-ELISA on μPADs.

Capillary Flow-Driven Microfluidics Combined with a Paper Device for Fast User-Friendly Detection of Heavy Metals in Water.

Human exposure to heavy metals is a concerning global problem because of its detrimental effect on our health and ecosystem. Assessing the levels of these metals is cost- and labor-intensive and nonuser friendly because current analysis approaches typically rely on heavy instrumentations like inductively coupled plasma-mass spectrometry, which is only possible in centralized labs. Hence, simple economical detection methods are in high demand in developing countries and areas with insufficient infrastructure, professional experts, and appropriate environmental treatment. Several microfluidic paper-based analytical devices have been reported as promising alternatives to conventional testing methods for on-site heavy metal detection. Paper-based microfluidics are advantageous because of their simple fabrication, biodegradability, low cost, and ability to operate without pumps. However, typical assay times for current platforms are slow, and they typically rely on pipetting a fixed volume into the assay cards. This adds complexity in actual field scenarios. Here, we report a novel, inexpensive, and straightforward capillary-driven microfluidic device combined with paper for rapid and user-friendly detection of Ni(II), Cu(II), and Fe(III) in water. A colorimetric approach was adopted to quantify these metals. The device was able to produce a homogeneous color signal within 8 s of sample insertion. The limit of detection and limit of quantification were calculated to be 2 and 6.67 ppm for nickel, 0.3 and 1 ppm for Cu, and 1.1 and 3.67 ppm for Fe, respectively. The majority (>90%) of the collected samples showed recovery in the 80-110% range with acceptable accuracy and precision (<15% RSD) for a colorimetric device. This technique can be beneficial for rapidly assessing heavy metal exposure in drinking and surface water at drastically reduced assay time and is the first example of capillary flow-driven microfluidic devices as a transport medium for heavy metal detection.

Hollow microneedle microfluidic paper-based chip for biomolecules rapid sampling and detection in interstitial fluid

The interstitial fluid (ISF) contains rich bioinformation for disease diagnosis and healthcare monitoring. However, the efficient sampling and detection of the biomolecules in ISF is still challenging. Herein, we develop a facile but versatile ISF analysis platform by combining controllable hollow microneedles (HMNs) and elaborate microfluidic paper-based analytical devices (μPADs). The HMNs and μPADs was fixed in a bottom PDMS layer. A top PDMS layer containing a cylindrical cavity to produce negative pressure for sampling was packaged on the bottom PDMS layer. The HMNs enable efficient and swift sampling of sufficient ISF to the μPADs through one-touch finger operation without extra manipulations. The μPADs realized to simultaneously detect glucose and lactic acid in the detection area to produce chromogenic agents and analyzed by the self-programed RGB application (APP) in smartphones. The HMN microfluidic paper-based chip provides a point-of-care platform for accurate detection of biomolecules in ISF, holding great promise in the development of wearable device.

Biomarker Detection in Early Diagnosis of Cancer: Recent Achievements in Point-of-Care Devices Based on Paper Microfluidics

Microfluidics is very crucial in lab-on-a-chip systems for carrying out operations in a large-scale laboratory environment on a single chip. Microfluidic systems are miniaturized devices in which the fluid behavior and control can be manipulated on a small platform, with surface forces on the platform being greater than volumetric forces depending on the test method used. In recent years, paper-based microfluidic analytical devices (μPADs) have been developed to be used in point-of-care (POC) technologies. μPADs have numerous advantages, including ease of use, low cost, capillary action liquid transfer without the need for power, the ability to store reagents in active form in the fiber network, and the capability to perform multiple tests using various measurement techniques. These benefits are critical in the advancement of paper-based microfluidics in the fields of disease diagnosis, drug application, and environment and food safety. Cancer is one of the most critical diseases for early detection all around the world. Detecting cancer-specific biomarkers provides significant data for both early diagnosis and controlling the disease progression. μPADs for cancer biomarker detection hold great promise for improving cure rates, quality of life, and minimizing treatment costs. Although various types of bioanalytical platforms are available for the detection of cancer biomarkers, there are limited studies and critical reviews on paper-based microfluidic platforms in the literature. Hence, this article aims to draw attention to these gaps in the literature as well as the features that future platforms should have.

Fabrication of microfluidic paper-based channels by inkjet printing process for analytical applications

Microfluidic paper-based channels play an important role in microfluidic paper-based analytical devices (μPADs). There are some fabrication methods which could be utilised to fabricate microfluidic channels on paper substrate. Among these methods, inkjet printing process is considered as a promising fabrication method with many advantages such as low-cost, material saving, high precision, etc. The aim of this work is to apply inkjet printing technology to fabricate paper channels of μPADs. A new design of μPAD was proposed in this paper to demonstrate how to fabricate inkjet-printed hydrophobic lines to make paper-based biosensor. Biological target of our μPADs is human chOrionic gonadotropin (hCG). Colorimetric signals from μPADs were captured by digital camera and measured by ImageJ software, which showed that these μPADs can determine hCG in the range from 1,000 to 10,000 ng ml−1. These results showed that piezoelectric inkjet printing technology can fabricate 250 μm-width hydrophobic lines on paper substrate, helping in fabricating μPADs in next applications.