Fluid Flow Dynamics in Partially Saturated Paper.

This study presents an integrated approach to understanding fluid dynamics in Microfluidic Paper-Based Analytical Devices (µPADs), combining empirical investigations with advanced numerical modeling. Paper-based devices are recognized for their low cost, portability, and simplicity and are increasingly applied in health, environmental monitoring, and food quality analysis. However, challenges such as lack of flow control and the need for advanced detection methods have limited their widespread adoption. To address these challenges, our study introduces a novel numerical model that incorporates factors such as pore size, fiber orientation, and porosity, thus providing a comprehensive understanding of fluid dynamics across various saturation levels of paper. Empirical results focused on observing the wetted length in saturated paper substrates. The numerical model, integrating the Highly Simplified Marker and Cell (HSMAC) method and the High Order accuracy scheme Reducing Numerical Error Terms (HORNET) scheme, successfully predicts fluid flow in scenarios challenging for empirical observation, especially at high saturation levels. The model effectively mimicked the Lucas-Washburn relation for dry paper and demonstrated the increasing time requirement for fluid movement with rising saturation levels. It also accurately predicted faster fluid flow in Whatman Grade 4 filter paper compared with Grade 41 due to its larger pore size and forecasted an increased flow rate in the machine direction fiber orientation of Whatman Grade 4. These findings have significant implications for the design and application of µPADs, emphasizing the need for precise control of fluid flow and the consideration of substrate microstructural properties. The study's combination of empirical data and advanced numerical modeling marks a considerable advancement in paper-based microfluidics, offering robust frameworks for future development and optimization of paper-based assays.

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.

Paper-Based Bi-Material Cantilever Actuator Bending Behavior and Modeling.

In this paper, the behavior of the Bi-Material Cantilever (B-MaC) response deflection upon fluidic loading was experimentally studied and modeled for bilayer strips. A B-MaC consists of a strip of paper adhered to a strip of tape. When fluid is introduced, the paper expands while the tape does not, which causes the structure to bend due to strain mismatch, similar to the thermal loading of bi-metal thermostats. The main novelty of the paper-based bilayer cantilevers is the mechanical properties of two different types of material layers, a top layer of sensing paper and a bottom layer of actuating tape, to create a structure that can respond to moisture changes. When the sensing layer absorbs moisture, it causes the bilayer cantilever to bend or curl due to the differential swelling between the two layers. The portion of the paper strip that gets wet forms an arc, and as the fluid advances and fully wets the B-MaC, the entire B-MaC assumes the shape of the initial arc. This study showed that paper with higher hygroscopic expansion forms an arc with a smaller radius of curvature, whereas thicker tape with a higher Young's modulus forms an arc with a larger radius of curvature. The results showed that the theoretical modeling could accurately predict the behavior of the bilayer strips. The significance of paper-based bilayer cantilevers lies in their potential applications in various fields, such as biomedicine, and environmental monitoring. In summary, the novelty and significance of paper-based bilayer cantilevers lie in their unique combination of sensing and actuating capabilities using a low-cost and environmentally friendly material.

Development of a New Lab-on-Paper Microfluidics Platform Using Bi-Material Cantilever Actuators for ELISA on Paper.

In this paper, we present a novel and cost-effective lab-on-paper microfluidics platform for performing ELISA autonomously, with no user intervention beyond adding the sample. The platform utilizes two Bi-Material Cantilever Valves placed in a specially designed housing. The integration of these valves in a specific channel network forms a complete fluidic logic circuit for performing ELISA on paper. The housing also incorporates an innovative reagent storage and release mechanism that minimizes variability in the volume of reagents released into the reagent pads. The platform design was optimized to minimize variance in the time of fluid wicking from the reagent pad, using a randomized design of experiment. The platform adheres to the World Health Organization's ASSURED principles. The optimized design was used to conduct an ELISA for detecting rabbit immunoglobulin G (IgG) in a buffer, with a limit of detection of 2.27 ng/mL and a limit of quantification of 8.33 ng/mL. This represents a 58% improvement over previous ELISA methods for detecting rabbit IgG in buffer using portable microfluidic technology.

A Parametric Study on a Paper-Based Bi-Material Cantilever Valve.

The novel paper-based Bi-Material Cantilever (B-MaC) valve allows the autonomous loading and control of multiple fluid reagents which contributes to the accurate operation of paper-based microfluidic devices utilized for biological and chemical sensing applications. In this paper, an extensive parametric study is presented to evaluate the effects of key geometric parameters of the valve, such as paper direction, cantilever width, paper type, tape type, and sample volume, in addition to the effects of relative humidity and temperature on the functionality of the B-MaC and to provide a better understanding of the rate of fluid flow and resulting deflection of the cantilever. Machine direction, cantilever width, paper type, and tape type were found to be important parameters that affect the B-MAC's activation time. It was also observed that the rate of fluid imbibition in the B-MaC is considerably affected by change in humidity for high (55 °C) and low (25 °C) temperatures, while humidity levels have no significant effect during imbibition in the B-MaC at an ambient temperature of 45 °C. It was also found that a minimum distance of 4 mm is required between the B-MaC and the stationary component to prevent accidental activation of the B-MaC prior to sample insertion when relative humidity is higher than 90% and temperature is lower than 35 °C. The rate of fluid imbibition that determines the wetted length of the B-MaC and the final deflection of the cantilever are critical in designing and fabricating point-of-care microfluidic paper-based devices. The B-MaC valve can be utilized in a fluidic circuit to sequentially load several reagents, in addition to the sample to the detection area.

A Colorimetric Dip Strip Assay for Detection of Low Concentrations of Phosphate in Seawater.

Nutrient pollution remains one of the greatest threats to water quality and imposes numerous public health and ecological concerns. Phosphate, the most common form of phosphorus, is one of the key nutrients necessary for plant growth. However, phosphate concentration in water should be carefully monitored for environmental protection requirements. Hence, an easy-to-use, field-deployable, and reliable device is needed to measure phosphate concentrations in the field. In this study, an inexpensive dip strip is developed for the detection of low concentrations of phosphate in water and seawater. In this device, ascorbic acid/antimony reagent was dried on blotting paper, which served as the detection zone, and was followed by a wet chemistry protocol using the molybdenum method. Ammonium molybdate and sulfuric acid were separately stored in liquid form to significantly improve the lifetime of the device and enhance the reproducibility of its performance. The device was tested with deionized water and Sargasso Sea seawater. The limits of detection and quantification for the optimized device using a desktop scanner were 0.134 ppm and 0.472 ppm for phosphate in water and 0.438 ppm and 1.961 ppm in seawater, respectively. The use of the portable infrared lightbox previously developed at our lab improved the limits of detection and quantification by a factor of three and were 0.156 ppm and 0.769 ppm for the Sargasso Sea seawater. The device's shelf life, storage conditions, and limit of detection are superior to what was previously reported for the paper-based phosphate detection devices.

A New Paper-Based Microfluidic Device for Improved Detection of Nitrate in Water.

In this paper, we report a simple and inexpensive paper-based microfluidic device for detecting nitrate in water. This device incorporates two recent developments in paper-based technology suitable for nitrate detection and has an optimized microfluidic design. The first technical advancement employed is an innovative fibrous composite material made up of cotton fibers and zinc microparticles that can be incorporated in paper-based devices and results in better nitrate reduction. The second is a detection zone with an immobilized reagent that allows the passage of a larger sample volume. Different acids were tested-citric and phosphoric acids gave better results than hydrochloric acid since this acid evaporates completely without leaving any residue behind on paper. Different microfluidic designs that utilize various fluid control technologies were investigated and a design with a folding detection zone was chosen and optimized to improve the uniformity of the signal produced. The optimized design allowed the device to achieve a limit of detection and quantification of 0.53 ppm and 1.18 ppm, respectively, for nitrate in water. This accounted for more than a 40% improvement on what has been previously realized for the detection of nitrate in water using paper-based technology.

A new paper-based platform technology for point-of-care diagnostics.

Currently, the Lateral flow Immunoassays (LFIAs) are not able to perform complex multi-step immunodetection tests because of their inability to introduce multiple reagents in a controlled manner to the detection area autonomously. In this research, a point-of-care (POC) paper-based lateral flow immunosensor was developed incorporating a novel microfluidic valve technology. Layers of paper and tape were used to create a three-dimensional structure to form the fluidic network. Unlike the existing LFIAs, multiple directional valves are embedded in the test strip layers to control the order and the timing of mixing for the sample and multiple reagents. In this paper, we report a four-valve device which autonomously directs three different fluids to flow sequentially over the detection area. As proof of concept, a three-step alkaline phosphatase based Enzyme-Linked ImmunoSorbent Assay (ELISA) protocol with Rabbit IgG as the model analyte was conducted to prove the suitability of the device for immunoassays. The detection limit of about 4.8 fm was obtained.

A fluidic diode, valves, and a sequential-loading circuit fabricated on layered paper.

Current microfluidic paper-based devices lack crucial components for fluid manipulation. We created a fluidic diode fabricated entirely on a single layer of paper to control the wicking of fluids. The fluidic diode is a two-terminal component that promotes or stops wicking along a paper channel. We further constructed a trigger and a delay valve based on the fluidic diode. Furthermore, we demonstrated a high-level functional circuit, consisting of a diode and a delay valve, to manipulate two fluids in a sequential manner. Our study provides new, transformative tools to manipulate fluid in microfluidic paper-based devices.

A blocking-free microfluidic fluorescence heterogeneous immunoassay for point-of-care diagnostics.

In this article, a rapid, sensitive, and disposable microfluidic immunosensor is presented for point-of-care (POC) testing and clinical diagnosis. For the first time, the blocking process is eliminated from a microfluidic heterogeneous immunoassay by using protein A functionalized polydimethylsiloxane microchannels. The nonspecific binding of the assay is maintained around the chip background level by using a pair of antibodies with different affinity to protein A under optimized experimental conditions. C-reactive protein (CRP), a biomarker for inflammation and cardiovascular disease risk assessment, is selected as a model analyte to demonstrate the sensitivity of this blocking-free microfluidic heterogeneous immunoassay. A four parameter logistic function is used to model and assess the data. The limit of detection obtained is 0.54 µg/mL, which is lower than the cut-off value for clinical diagnosis. The overall assay is completed in 5 min. The protein A modified PDMS chips wet-stored at 4°C can maintain biofunctionality up to 14 months. The developed blocking-free microfluidic heterogeneous immunoassay will immediately provide benefits to most immunosensing microdevices targeted for POC diagnostics by shortening analysis time, simplifying fluid transportation, reducing sample consumption, and lowering waste generation.