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Cell-free expression systems have drawn increasing attention as a tool to achieve complex biological functions outside of the cell. Several applications of the technology involve the delivery of functionality to challenging environments, such as field-forward diagnostics or point-of-need manufacturing of pharmaceuticals. To achieve these goals, cell-free reaction components are preserved using encapsulation or lyophilization methods, both of which often involve an embedding of components in porous matrices like paper or hydrogels. Previous work has shown a range of impacts of porous materials on cell-free expression reactions. Here, we explored a panel of 32 paperlike materials and 5 hydrogel materials for the impact on reaction performance. The screen included a tolerance to lyophilization for reaction systems based on both cell lysates and purified expression components. For paperlike materials, we found that (1) materials based on synthetic polymers were mostly incompatible with cell-free expression, (2) lysate-based reactions were largely insensitive to the matrix for cellulosic and microfiber materials, and (3) purified systems had an improved performance when lyophilized in cellulosic but not microfiber matrices. The impact of hydrogel materials ranged from completely inhibitory to a slight enhancement. The exploration of modulating the rehydration volume of lyophilized reactions yielded reaction speed increases using an enzymatic colorimetric reporter of up to twofold with an optimal ratio of 2:1 lyophilized reaction to rehydration volume for the lysate system and 1.5:1 for the purified system. The effect was independent of the matrices assessed. Testing with a fluorescent nonenzymatic reporter and no matrix showed similar improvements in both yields and reaction speeds for the lysate system and yields but not reaction speeds for the purified system. We finally used these observations to show an improved performance of two sensors that span reaction types, matrix, and reporters. In total, these results should enhance efforts to develop field-forward applications of cell-free expression systems.

A wet‐vacuum‐based collection method with the M‐Vac® was compared to a wet‐swabbing collection method by examining the recovery of diluted blood on 22 substrates of varying porosity. The wet‐vacuum method yielded more total nuclear DNA than wet‐swabbing on 18 porous substrates, recovering on average 12 times more DNA. However, both methods yielded comparable amounts of total DNA on two porous and two nonporous substrates. In no instance did wet‐swabbing significantly recover more DNA. The wet‐vacuum method also successfully collected additional DNA on previously swabbed substrates. Mitochondrial DNA yields were assessed, and outcomes were generally similar to the nuclear DNA outcomes described above. Results demonstrate that wet‐vacuuming may serve as an alternative collection method to swabbing on difficult porous substrates and could potentially recover additional DNA on previously swabbed substrates. However, swabbing remains the preferred collection method on substrates with visible stains and/or nonporous surfaces for reasons of convenience, simplicity, and lower cost relative to the wet‐vacuum method.

A complete mathematical model for electromigration in paper-based analytical devices is derived, based on differential equations describing the motion of fluids by pressure sources and EOF, the transport of charged chemical species, and the electric potential distribution. The porous medium created by the cellulose fibers is considered like a network of tortuous capillaries and represented by macroscopic parameters following an effective medium approach. The equations are obtained starting from their open-channel counterparts, applying scaling laws and, where necessary, including additional terms. With this approach, effective parameters are derived, describing diffusion, mobility, and conductivity for porous media. While the foundations of these phenomena can be found in previous reports, here, all the contributions are analyzed systematically and provided in a comprehensive way. Moreover, a novel electrophoretically driven dispersive transport mechanism in porous materials is proposed. Results of the numerical implementation of the mathematical model are compared with experimental data, showing good agreement and supporting the validity of the proposed model. Finally, the model succeeds in simulating a challenging case of free-flow electrophoresis in paper, involving capillary flow and electrophoretic transport developed in a 2D geometry. Fil: Schaumburg, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina Fil: Kler, Pablo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Centro de Investigaciones en Métodos Computacionales. Universidad Nacional del Litoral. Centro de Investigaciones en Métodos Computacionales; Argentina Fil: Berli, Claudio Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina

In previous work, our group discovered a phenomenon in which a mixed polymer-salt or mixed micellar aqueous two-phase system (ATPS) separates into its two constituent phases as it flows within paper. While these ATPSs worked well in their respective studies to concentrate the target biomarker and improve the sensitivity of the lateral-flow immunoassay, different ATPSs can be advantageous for new applications based on factors such as biomarker partitioning or biochemical compatibility between ATPS and sample components. However, since the mechanism of phase separation in porous media is not completely understood, introducing other ATPSs to paper is an unpredictable process that relies on trial and error experiments. This is especially true for polymer-polymer ATPSs in which the characteristics of the two phases appear quite similar. Therefore, our group aimed to develop semiquantitative guidelines for choosing ATPSs that can phase separate in paper. In this work, we evaluated the Washburn equation and its parameters as a potential mathematical framework to describe the flow behavior of polymer-salt and micellar ATPSs in fiberglass paper. We compared bulk phase fluid characteristics and identified the viscosity difference between the phases as a key determinant of the potential for phase separation in paper. We then used this parameter to predict the phase separation capabilities of polyethylene glycol (PEG)-dextran ATPSs in paper and control the composition of the leading and lagging phases. We also, for the first time, successfully demonstrated the phase separation phenomenon in hydrogels, thereby extending its application and potential benefits to an alternative porous medium.

Porous magnesium oxide (MgO) films on carbon fiber paper (CF) have been successfully fabricated in a hydrothermal route at different calcination temperatures. The CF@MgO samples (CF@MgO-300, -400, and -500) show different morphologies with the increasing surface area from 3 for CF to 27 m2 g−1 for CF@MgO-400. Among the four investigated samples, the CF@MgO-400 exhibits the highest phosphate removal ability (~ 1230 mg g−1) with promising applications for the large-scale utilization at low cost.

Microfluidic paper-based analytical devices (μPADs) are simple but powerful analytical tools that are gaining significant recent attention due to their many advantages over more traditional monitoring tools. These include being inexpensive, portable, pump-free, and having the ability to store reagents. One major limitation of these devices is slow flow rates, which are controlled by capillary action in the hydrophilic pores of cellulosic paper. Recent investigations have advanced the flow rates in μPADs through the generation of a gap or channel between two closely spaced paper sheets. This multilayered format has opened up μPADs to new applications and detection schemes, where large gap sizes (>300 μm) provide at least 169× faster flow rates than single-layer μPADs, but do not conform to established mathematical models for fluid transport in porous materials, such as the classic Lucas-Washburn equation. In the present study, experimental investigations and analytical modeling are applied to elucidate the driving forces behind the rapid flow rates in these devices. We investigate a range of hypotheses for the systems fluid dynamics and establish a theoretical model to predict the flow rate in multilayered μPADs that takes into account viscous dissipation within the paper. Device orientation, sample addition method, and the gap height are found to be critical concerns when modeling the imbibition in multilayered devices.

International audience; The Covid-19 coronavirus, SARS-CoV-2, is inactivated much faster on paper (3 h) than on plastic (7 d). By classifying materials according to virus stability on their surface, the following list is obtained (from long to short stability): polypropylene (mask), plastic, glass, stainless steel, pig skin, cardboard, banknote, cotton, wood, paper, tissue, copper. These observations and other studies suggest that SARS-CoV-2 may be inactivated by dryness on water absorbent porous materials but sheltered by long-persisting micro-droplets of water on waterproof surfaces. If such physical phenomenons were confirmed by direct evidence, the persistence of the virus on any surface could be predicted, and new porous objects could be designed to eliminate the virus faster.

In this study we investigated the underlying mechanisms of coffee stain formation on paper - a porous substrate. When a drop of aqueous liquid containing dispersed materials dries on paper, the faster water evaporation rate at the edge of the drop drives the redistribution of the dispersed materials to the edge of the drop-covered area on paper. Deegan's discovery has been accepted as the "coffee ring" phenomenon, which has wide implications. However, stains left by dried aqueous liquids on paper are not always ring-shaped. This is because, besides Deegan's coffee ring effect, the transport of dispersed materials in porous media is affected by chromatographic and filtration effects, which are highly dependent upon the properties of the substrates. In this work we clarify the influences of different mechanisms to the dispersed materials redistribution in paper. Refereed/Peer-reviewed

A novel washing technique for microfluidic paper-based analytical devices (μPADs) that is based on the spontaneous capillary action of paper and eliminates unbound antigen and antibody in a sandwich immunoassay is reported. Liquids can flow through a porous medium (such as paper) in the absence of external pressure as a result of capillary action. Uniform results were achieved when washing a paper substrate in a PDMS holder which was integrated with a cartridge absorber acting as a porous medium. Our study demonstrated that applying this washing technique would allow μPADs to become the least expensive microfluidic device platform with high reproducibility and sensitivity. In a model μPAD assay that utilized this novel washing technique, C-reactive protein (CRP) was detected with a limit of detection (LOD) of 5 μg mL−1.

International audience; The aim of this work is the experimental determination of effective transport properties of porous media consisting of compacted pieces of cardboard and polyethylene (PE). The proposed method itself is more general and can be applied to many different materials and contexts. Three major transport properties were determined: porosity, tortuosity factor and permeability. Three parameters characterizing the media were varied over a wide range: the bulk density, the size of the elements entering the mix, and the proportion of cardboard and PE in the mix. The properties were measured by means of a specially designed experimental device based on miscible gas tracing. The porosity and tortuosity factor were simultaneously determined by parametric identification, based on the experimental sample output response to an inlet gas concentration step change compared to the results of a direct numerical model. Permeability was calculated in the standard way from the measurement of the pressure drop across the sample. The reproducibility of the measurements was very good. It was found that changing the material density of the medium significantly affects all three structural properties. When the bulk density is varied between 300 and 900 kg m À3 , the tortuosity factor varies in a range as large as 18-8 and the permeability decreases by a ratio of 2-3. The tortuosity factor shows unusual variation, characterized by a decrease when density is increased above 500 kg m À3. The size of the elements does not significantly affect the structural properties of the medium in the range of parameters studied.

Flow monitoring in porous materials is critical for the engineering of paper-based microfluidic bioassays. Here, we present an electrochemical-sensor system that monitors the liquid flow in porous materials without affecting the real flow in paper-strip samples. The developed microfluidic sensor records an amperometric signal created by the solution movement mediated by paper wicking. This approach allows the in situ monitoring of the different hydrodynamic conditions of a specific paper geometry or composition. In addition, the method proposed in this work was employed to characterise the fluid flow of different nitrocellulose paper strips after oxygen-plasma treatment or dextran coating. The dextran fluid-flow modifiers were further used on the paper strip-based assays as means of signal enhancement. The proposed electrochemical-sensing method offers a valuable alternative to existing optical-based monitoring techniques for flow measurement in paper-based microfluidic systems.

Theoretical and experimental study of the paper drying process are presented. A mathematical model developed for combined heat ad mass transfer analysis of paper drying is given for both impinging air jets and through air drying methods. In this model, it is simply assumed that during the drying period of the paper has porous media and on the drying surface the vapour pressure of the evaporating liquid remains at a quasi-saturated value corresponding to the temperature of the liquid. The calculated transient paper temperatures in both methods agree well with the experimental results.

In this work, we propose a new approach to modeling multiphase flow and solute transport through a stack of thin porous layers. Currently, numerical simulation of thin layers involves discretization across the layer thickness. In our new approach, thin porous layers are treated as a bunch of two-dimensional (2D) interacting continua. Macroscale balance laws are formulated in terms of thickness-averaged material properties. A number of exchange terms are employed to account for exchanges of mass, momentum, energy, and entropy between two neighboring layers. The entropy inequality is then exploited for obtaining constitutive equations to close the problem under study. As an example, simplified governing equations are derived for a system of air–water flow and heat transfer through two thin porous layers. In comparison to previous macroscale models, our model possesses the following distinctive advantages: (1) it is rigorous thermodynamics-based model; (2) it is formulated in terms of layer-thickness-averaged material properties which are usually easily measurable; and (3) it reduces 3D modeling to 2D leading to a very significant reduction of computational efforts.

Models of reactive uptake of ozone in indoor environments generally describe materials through aerial (horizontal) projections of surface area, a potentially limiting assumption for porous materials. We investigated the effect of changing porosity/pore size, material thickness, and chamber fluid mechanic conditions on the reactive uptake of ozone to five materials: two cellulose filter papers, two cementitious materials, and an activated carbon cloth. Results include (1) material porosity and pore size distributions, (2) effective diffusion coefficients for ozone in materials, and (3) material-ozone deposition velocities and reaction probabilities. At small length scales (0.02-0.16 cm) increasing thickness caused increases in estimated reaction probabilities from 1 × 10(-6) to 5 × 10(-6) for one type of filter paper and from 1 × 10(-6) to 1 × 10(-5) for a second type of filter paper, an effect not observed for materials tested at larger thicknesses. For high porosity materials, increasing chamber transport-limited deposition velocities resulted in increases in reaction probabilities by factors of 1.4-2.0. The impact of physical properties and transport effects on values of the Thiele modulus, ranging across all materials from 0.03 to 13, is discussed in terms of the challenges in estimating reaction probabilities to porous materials in scenarios relevant to indoor environments.

Novel methods are demonstrated that enable controlled spatial and temporal rehydration of dried reagents in a porous matrix. These methods can be used in paper-based microfluidic assays to define reagent concentrations over time at zones downstream for improved performance, and can reduce costs by simplifying the manufacturing process with the use of a single porous substrate. First, the creation of uniform reagent pulses from patterned arrays of dried reagent is demonstrated. Second, reagents are stored dry in separate regions of the porous matrix so that they can be combined upon rehydration for immediate use in the device. Third, reagents are reconstituted sequentially from dry storage depots with tunable delivery times. Fourth, the total time for dissolution is varied to achieve a range of reagent delivery times to a downstream region. Finally, the utility of these control methods is demonstrated in the context of real-time reagent rehydration and mixing on a porous device.

Activated carbon with large specific surface area and well-developed porosity was prepared from pyrolysis of K2CO3-impregnated lignin precipitated from reed pulp black liquors. The impregnation ratio was 1:1. The effect of activation temperature upon the Brunauer-Emmett-Teller (BET) specific surface area and pore volume of the carbon was closely investigated. Increasing activation temperature led to an opening and widening of the porous structure below 800'C. Above 800'C, the excess widening of pore led to the decrease of BET surface area and micropore volume. The BET surface area and pore volume of the carbon activated at 800 degrees C were 1395 m(2) g(-1) and 0.7702 ml g(-1) , respectively. The potential application of the carbon activated at 800 degrees C for removal of Cr (VI) was also investigated. The experimental results showed that it had good adsorption capacity.