Paper-based Devices Research Articles

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.

Paper-based uric acid assay in whole blood samples by Zn2+ protein precipitation and enzyme-free colorimetric detection.

Uric acid (UA) is an important biomarker, as a high concentration in blood can lead to gout and further renal syndrome. Although several point-of-caretesting (POCT) devices have been reported to detect UA, there are some limitations such as the requirement for uricase and the complicated pretreatment of serum/plasma samples, which restricts their use at home or in undeveloped areas. In this work, we developed an approach by applying Zn2+ to precipitate proteins and cells in whole blood to avoid interference with the chromogenic reaction. We used carboxymethylcellulose (CMC) to immobilize tetramethylbenzidine (TMB) on a nitrocellulose membrane for colorimetric detection. Using the oxidization properties of H2O2, which turns TMB into oxidized tetramethylbenzidine (TMBox) in the presence of catalyst gold nanoparticles (AuNPs), we successfully constructed an enzyme-free paper-based POCT device using the reduction reaction of UA and TMBox for simple, speedy, and cheap colorimetric detection of UA, achieving a detection time of 8min, a linear range of 0-150μg/mL, and an LOD of 25.79μg/mL. The UA concentration in whole blood samples was further measured and correlated well with the clinical value (R2 = 0.8212). Thus, the proposed assay has the potential for POCT diagnosis, monitoring, and prognosis of diseases related to UA.

Development and validation of spectrophotometric method and paper-based microfluidic devices for the quantitative determination of Amoxicillin in pure form and pharmaceutical formulations

There is a growing need for easy-to-use, low cost and portable quantitative assays to determine active pharmaceutical ingredients in the pharmaceutical industry. Here, we developed a batch spectrophotometric method and a method employing a paper-based microfluidic device for the estimation of Amoxicillin (AMX) in pure solution and pharmaceutical preparations. The detection depends on the coupling reaction of Amoxicillin with diazotized sulfadimidine (DSDM) in an alkaline medium. The yellow azo dye reaction product was measured at λmax 425 nm and linearity was observed from 2 to 30 mg L−1 with a detection limit of 0.32 mg L−1 and a quantification limit of 1.2 mg L−1 was found. The reaction was then transferred onto the paper-based microfluidic device and a plateau change in color intensity was found above 10 mg L−1. Thus, the paper-based microfluidic device can be applied for the semi-quantitative determination of Amoxicillin in pure solution and commercial pharmaceutical products for rapid screening.

Thermally programmable time delay switches for multi-step assays in paper-based microfluidics

Paper-based microfluidic devices offer advantages such as low cost and disposability for point-of-care diagnostic applications. However, actuation of fluids on paper can be a challenge in multi-step and complex assays. In this work, a thermally programmable time-delay switch (TPTDS) is presented which operates by causing delays in the fluid path of a microfluidics paper-based analytical device (μPAD) by utilizing screen-printed wax micro-bridges. The time-delay is achieved through an electrical power feedback loop which indirectly adjusts the temperature of each individual micro-bridge, melting the wax into the paper. The melted wax manipulates the fluid flow depending on its penetration depth into the paper channel, which is a function of the applied temperature. To demonstrate functionality of the proposed method, the TPTDS is employed to automate and perform the nitrate assay which requires sequential delivery of reagents. Colorimetric detection is used to quantify the results by utilizing an electronic color sensor.

High-Throughput Continuous Free-Flow Dielectrophoretic Trapping of Micron-Scale Particles and Cells in Paper Using Localized Nonuniform Pore-Scale-Generated Paper-Based Electric Field Gradients.

Dielectrophoresis (DEP) utilizes a spatially varying nonuniform electrical field to induce forces on suspended polarizable soft matter including particles and cells. Such nonuniformities are conventionally created using 2D or 3D micrometer-scale electrode arrays. Alternatively, insulator-based dielectrophoresis (iDEP) uses small micrometer-scale insulating structures to spatially distort and generate regions of localized field gradients to selectively trap, isolate, and concentrate bioparticles, including bacteria, viruses, red blood cells, and cancer cells from a suspending electrolyte solution. Despite significant advances in the microfabrication technology, the commercial adoption of DEP devices for soft matter manipulation remains elusive. One reason for low market penetration is a lack of low-cost and scalable fabrication methods to quickly microfabricate field-deforming structures to generate localized DEP-inducing electric field gradients. We propose here that paper-based devices can offer a low-cost and easy-to-use alternative to traditional iDEP devices. In this article, we demonstrate for the first time the ability to perform iDEP-style particle trapping using the naturally occurring micrometer-scale insulating porous structures of paper. In particular, we use polymeric laminated nonwoven fiberglass paper channels as a source of insulating structures for iDEP. We apply a flow of polarizable microparticles directly within the nonwoven channel and simultaneously drop an electric field perpendicular to the flow direction to induce DEP. We show the ability to readily trap and concentrate particles in paper by DEP with an applied voltage as low as 2 V using two different flow mechanisms: a constant fluid flow rate using an external pump and passive fluid flow by capillary wicking. Using a combination of micro computed tomography and finite element analysis, we then present a computational model to probe the microscale DEP force formation dynamics within the paper structure. This new paper-based iDEP platform enables the development of robust, low-cost, and portable next-generation iDEP systems for a wide variety of sample purification and liquid handling applications.

Instrument-free and rapid Vibrio parahaemolyticus biosensing based on coupled dual-signal of paper-needle and LnMOF

At present, most reported studies on paper-based sensing devices for food-borne pathogens usually rely on a single signal as the basis for judging the results, accompanied with inaccurate detection results. In this study, we used the lanthanide MOFs being covalently modified with aptamers (Apt-LnMOFs) as the sensing and recognition element of Vibrio parahaemolyticus (VP), and the paper-based needle shape device (paper-needle) as the visual signal output carrier. Finally, an accurate detection strategy for VP was successfully constructed. In terms of signal output, the two signal styles of Apt-LnMOFs diffusion distance on the paper-needle and R value (measured by the RGB analysis software of smartphone) are coupled to jointly improve the accuracy of detection results. The method is simple and fast, and the detection accuracy in real samples is 91%. This strategy is very suitable for on-site rapid detection of pathogens, and does not require large professional instruments.

Origami Paper-Based Electrochemical Immunosensor with Carbon Nanohorns-Decorated Nanoporous Gold for Zearalenone Detection

Nowadays, mycotoxin contamination in cereals and wastewater exposes a safety hazard to consumer health. This work describes the design of a simple, low-cost, and sensitive origami microfluidic paper-based device using electrochemical detection for zearalenone determination. The microfluidic immunosensor was designed on a paper platform by a wax printing process. The graphitized carbon working electrode modified with carbon nanohorns-decorated nanoporous gold showed a higher surface area, sensitivity, and adequate analytical performance. Electrodes were characterized by scanning electron microscopy, energy-dispersive spectroscopy, and cyclic voltammetry. The determination of zearalenone was carried out through a competitive immunoassay using specific antibodies immobilized by a covalent bond on the electrode surface. In the presence of HRP-labeled enzyme conjugate, substrate, and catechol, zearalenone was detected employing the developed immunosensor by applying −0.1 V to the working electrode vs silver as a pseudo-reference electrode. A calibration curve with a linear range between 10 and 1000 µg Kg−1 (R2 = 0.998) was obtained, and the limit of detection and quantification for the electrochemical immunosensor were 4.40 and 14.90 µg Kg−1, respectively. The coefficient of variation for intra- and inter-day assays was less than 5%. The selectivity and specificity of the sensor were evaluated, comparing the response against zearalenone metabolites and other mycotoxins that could affect the corn samples. Therefore, origami is a promising approach for paper-based electrochemical microfluidic sensors coupled to smartphones as a rapid and portable tool for in situ mycotoxins detection in real samples.

Quantification of creatinine in whole blood by a paper-based device using an RGB sensor.

Creatinine, an important biomarker for renal functions, is often conventionally estimated using the gold standard Jaffe reaction from blood, which involves the usage of a spectrophotometric measurement, thus restricting its utilization scope in point-of-care settings. Here, we report the development of a method for the single-step quantification of creatinine from whole blood using a paper-based microfluidic device. Our platform uses Whatman filter paper integrated with an LF1 membrane. The on-chip separation of blood plasma is achieved through the LF1 membrane, while the Whatman component of the device contains the embedded reagents for the Jaffe reaction. The combination of two different grades of paper enables a single-step quantification of creatinine as the separated blood plasma traverses to the reaction zone through capillary imbibition. Colorimetric readouts were quantified using an RGB sensor instead of a smartphone, which is highly platform dependent and incurs a relatively higher cost compared to the other components in typical point-of care (POC) devices. Our sensor was integrated within a 3D box, thereby making the detection virtually instrument free and perfectly suited for POC settings. The limit of detection (LOD) of our device was 0.219 mg dL-1, which falls within the lower range of physiological values. The coefficient of determination (R2) for the linearity and median accuracy were 0.978 and 94.047%, respectively. The relative standard deviation (RSD) for precision measurements remained below 5% for the developed protocol. Furthermore, we validated the performance of our device with 35 clinical samples in laboratory settings against the gold standard measurements. Our Bland-Altman plot as well as t-test and chi-square test results clearly confirmed the validity of our device within a 95% confidence interval.

Paper-based chemometer device for the estimation of α-amylase-a biomarker for pancreatitis.

Pancreatitis is a life-threatening inflammatory disease of the pancreas. In 2019, 34.8 out of 100 000 people suffered from acute pancreatitis globally. In humans, the level of α-amylase increases three times the normal value during pancreatitis. α-Amylase is an enzyme that hydrolyses α-1,4 glycosidic bonds of starch. In this study, we investigated a novel distance-based sensing method. We exploited the existing starch triiodide method, where the blue colour of starch-triiodide fades away and becomes colourless when α-amylase breaks the starch chain at the α-1-4 glycosidic bond. A hydrophilic channel was made on paper using a simple laser printer to create hydrophobic barriers. This channel was impregnated with starch triiodide, where α-amylase can turn it colourless. This distance covered by the change in colour is directly proportional to the concentration of α-amylase in a sample. Simulated samples with different concentrations of porcine α-amylase and pancreatin were used for testing using the developed paper-based chemometer device. The paper-based chemometer device was also tested with artificial blood serum with different concentrations of α-amylase. The R 2 of this device was found to be 0.9905, and the accuracy of the device when compared with a 2-chloro-4-nitrophenyl-α-d-maltotrioside method was found to be 95.54% with a sensitivity of 0.131 U L-1 mm-1. Correlation test also showed that the paper-based chemometer device for α-amylase can be used as a testing device for artificial blood serum. This is a preliminary investigation that shows promising results. The chemometer devices stored in air-tight packets at 4-8 °C in a refrigerator did not lose the colour intensity until day 90 and retained an accuracy of 94.5%. However, the device needs to be evaluated in clinical settings prior to using it for measuring α-amylase in patients.

Surface modification of paper-based microfluidic devices via initiated chemical vapor deposition.

Paper-based microfluidic devices offer an ideal platform for biological and environmental detection because they are low-cost, small, disposable, and fill by natural capillary action. In this tutorial review, we discuss the surface modification of paper-based microfluidic devices with functional polymers using the initiated chemical vapor deposition (iCVD) process. The iCVD process is solventless and therefore ideal for coating cellulose paper because there are no surface tension effects or solvent compatibility issues. The process can also be scaled up for roll-to-roll manufacturing. The chemical functionality of the iCVD coating can be tuned by varying the monomer and the structure of the coating can be tuned by varying the processing parameters.

A separation-free paper-based hydrogel device for one-step reactive oxygen species determination by a smartphone.

Paper-based analytical devices (PADs) are very convenient for determining biomarkers in point-of-care (POC) diagnosis while requiring sample pre-treatment or impurity separation. This study reports a novel hydrogel-coupled, paper-based analytical device (PAD) for separation-free H2O2 colorimetric detection in both aqueous solution and cell lysis with sample-to-answer analysis by directly loading into the sample test zone. By encapsulating an inorganic mimic enzyme and chromogenic substrate into the sodium alginate (SA) hydrogel, amplification of the color signal after catalyzing the substrate could be achieved. Taking advantage of the nanoscale porous structure of the hydrogel and the lateral flow channel of the PAD, large interference fragments or bio-macromolecules are prevented from diffusing into the chromogenic reaction, whereas the small target molecules enter the sensing region to trigger the catalytic reaction. This method demonstrated a rapid and accurate analysis with a limit of detection as low as 0.06 mM and detection selectivity. Our proposed device requires no enzyme and is separation-free, portable, easy-to-fabricate, and low-cost, and may offer a platform for quantitative or qualitative analysis of other analytes in body fluids for POC applications.

One-drop chemosensing of dapoxetine hydrochloride using opto-analysis by multi-channel μPAD decorated silver nanoparticles: introducing a paper-based microfluidic portable device/sensor toward naked-eye pharmaceutical analysis by lab-on-paper technology.

Dapoxetine (DPX) belongs to the selective serotonin reuptake inhibitor (SSRI) class and functions by blocking the serotonin transporter and increasing serotonin activity, thereby delaying ejaculation. Therefore, monitoring of the concentration of DPX in human biofluids is important for clinicians. In this study, application of silver nanoparticles with the morphology of prisms (AgNPrs) for the sensitive measurement of DPX using colorimetric chemosensing and the spectrophotometric method was investigated. Also, DPX was determined in real samples using the spectrophotometry method. Based on the obtained results, all of the detection process in colorimetric assay is related to morphological reform of AgNPrs after it's specific electrostatic and covalent interaction with DPX as analyte. The UV-vis results indicate that the proposed AgNPrs-based chemosensing system has a wide range of linearity (0.01 μM to 1 mM) with a low limit of quantification of 0.01 μM in human urine samples, which is suitable for clinical analysis of this drug in human urine samples. It is important to point out that, this chemosensing strategy showed inappropriate analytical results for the detection of DPX in human urine samples which is a novelty of this platform. Finally, the optimized microfluidic paper-based analytical device (μPAD) was integrated with the colorimetric analysis of DPX to provide a time/color system for estimating analyte concentration by a portable substrate toward in situ and on-site biomedical analysis. Interestingly, the analytical validation tests showed appropriate results with great stability, which may facilitate commercialization of the engineered substrate. For the first time, in order to provide a simple and portable colorimetric/spectrophotometric recognition system to sensitive determination of DPX, an optimized pump-less microfluidic paper-based colorimetric device (μPCD) was introduced and validated for the real-time biomedical analysis of this analyte. According to the obtained results, this alternative approach is suitable for therapeutic drug monitoring (TDM) and biomedical analysis by miniaturized and cost-beneficial devices.

A dual-mode homogeneous electrochemical-colorimetric biosensing sensor for carcinoembryonic antigen detection based on a microfluidic paper-based analysis device.

Dual-mode-based sensors have drawn a lot of interest due to their high accuracy and sensitivity compared to single-response systems. A simple electrochemical and colorimetric dual-mode sensor based on enzyme-linked immunosorbent assay (ELISA), without complex electrode surface modification, was developed for accurate and sensitive detection of carcinoembryonic antigen (CEA). The target CEA is recognized by an antibody coupled to horseradish peroxidase (HRP), which then oxidizes the substrate 3,3',5,5'-tetramethylbenzidine (TMB) to generate both a colorimetric and an electrochemical signal. A paper-based analysis device (μPAD) with dual-mode homogeneous sensing microfluidic was created; three paper-based detection areas for colorimetric testing, and a two-electrode embedded detection area for electrochemical testing. When applying colorimetric analysis technology, the linear range of CEA detection is 0.6-40 ng mL-1, and the limit of detection (LOD) is 0.2 ng mL-1. The linear range is 0.1-40 ng mL-1 and the LOD is 0.03 ng mL-1 by applying electrochemical analysis. The visibility and intuitiveness of colorimetry provide a reference for higher sensitivity and quick response of the electrochemical method. A smartphone application (APP) was also developed to realize the dual extraction of colorimetric signals. The colorimetric detection system based on ELISA can provide a new path for the development of electrochemical sensing and makes it have inherent self-verification and self-correction functions and is expected to provide more reliable and accurate detection results.

Screen printed 3D microfluidic paper-based and modifier-free electroanalytical device for clozapine sensing.

The increasing demand in healthcare for accessible and cost-effective analytical tools is driving the development of reliable platforms to the customization of therapy according to individual patient drug serum levels, e.g. of anti-psychotics in schizophrenia. A modifier-free microfluidic paper-based electroanalytical device (μPED) holds promise as a portable, sensitive, and affordable solution. While many studies focus on the working electrode catalysts, improvements by engineering aspects e.g. of the electrode arrangement are less reported. In our study, we demonstrate the enhanced capabilities of the 3D electrode layout of μPED compared to 2D μPED arrangements. We especially show that screen printing can be employed to prepare 3D μPEDs. We conducted a comparison of different 2D and 3D electrode arrangements utilizing cyclic voltammetry in [Fe(CN)6]3-/4-, along with square-wave voltammetry for clozapine (CLZ) sensing. Our findings reveal that the utilization of the 3D μPED leads to an increase in both the electrochemically active surface area and the electron transfer rate. Consequently, this enhancement contributes to improve sensitivity in the CLZ sensing. The 3D μPED clearly outperforms the 2D μPED arrangement in terms of signal strength. With the 3D μPED under the optimized conditions, a linear dose-response for a concentration range from 7.0 to 100 μM was achieved. The limit of detection and sensitivity was determined to be 1.47 μM and 1.69 μA μM-1 cm-2, respectively. This evaluation is conducted in the context of detection and determination of CLZ in a human blood serum sample. These findings underscore the potential of the 3D μPED for future applications in pharmacokinetic analyses and clinical tests to personalize the management of schizophrenia.

Vibration mixing for enhanced paper-based recombinase polymerase amplification.

Isothermal nucleic acid amplification tests (NAATs) are a vital tool for point-of-care (POC) diagnostics. These assays are well-suited for rapid, low-cost POC diagnostics for infectious diseases compared to traditional PCR tests conducted in central laboratories. There has been significant development of POC NAATs using paper-based diagnostic devices because they provide an affordable, user-friendly, and easy to store format; however, the difficulties in integrating separate liquid components, resuspending dried reagents, and achieving a low limit of detection hinder their use in commercial applications. Several studies report low assay efficiencies, poor detection output, and poorer limits of detection in porous membranes compared to traditional tube-based protocols. Recombinase polymerase amplification is a rapid, isothermal NAAT that is highly suited for POC applications, but requires viscous reaction conditions that has poor performance when amplifying in a porous paper membrane. In this work, we show that we can dramatically improve the performance of membrane-based recombinase polymerase amplification (RPA) of HIV-1 DNA and viral RNA by employing a coin cell-based vibration mixing platform. We achieve a limit of detection of 12 copies of DNA per reaction, nearly 50% reduction in time to threshold (from ∼10 minutes to ∼5 minutes), and an overall fluorescence output increase up to 16-fold when compared to unmixed experiments. This active mixing strategy enables reactions where the target and reaction cofactors are isolated from each other prior to the reaction. We also demonstrate amplification using a low-cost vibration motor for both temperature control and mixing, without the requirement of any additional heating components.

Unveiling the state of the art: a systematic review and meta-analysis of paper-based microfluidic devices.

This systematic review and meta-analysis present a comprehensive evaluation of paper-based microfluidic devices, focusing on their applications in immunoassays. These devices are emerging as innovative solutions to democratize access to diagnostic technologies, especially in resource-limited settings. Our review consolidates findings from diverse studies to outline advancements in paper-based microfluidic technology, including design intricacies and operational efficacy. Key advantages such as low cost, portability, and ease of use are highlighted. The review categorizes literature based on the design and operational nuances of these diagnostic tools, exploring various methodologies, fabrication techniques, detection methods, and applications, particularly in protein science. The meta-analysis extends to the diverse applications of these technologies, providing a framework for classifying and stratifying their uses in diagnostics. Notable findings include a critical analysis of performance metrics, such as sensitivity and specificity. The review addresses challenges, including the need for further validation and optimization for broader clinical applications. A critical discussion on the validation processes, including cross-validation and rigorous control testing, is provided to ensure the robustness of microfluidic devices. This study offers novel insights into the computational strategies underpinning these technologies and serves as a comprehensive roadmap for future research, potentially broadening the impact across the protein science universe.

Plasmonic silver and gold nanoparticles: shape- and structure-modulated plasmonic functionality for point-of-caring sensing, bio-imaging and medical therapy.

Silver and gold nanoparticles have found extensive biomedical applications due to their strong localized surface plasmon resonance (LSPR) and intriguing plasmonic properties. This review article focuses on the correlation among particle geometry, plasmon properties and biomedical applications. It discusses how particle shape and size are tailored via controllable synthetic approaches, and how plasmonic properties are tuned by particle shape and size, which are embodied by nanospheres, nanorods, nanocubes, nanocages, nanostars and core-shell composites. This article summarizes the design strategies for the use of silver and gold nanoparticles in plasmon-enhanced fluorescence, surface-enhanced Raman scattering (SERS), electroluminescence, and photoelectrochemistry. It especially discusses how to use plasmonic nanoparticles to construct optical probes including colorimetric, SERS and plasmonic fluorescence probes (labels/reporters). It also demonstrates the employment of Ag and Au nanoparticles in polymer- and paper-based microfluidic devices for point-of-care testing (POCT). In addition, this article highlights how to utilize plasmonic nanoparticles for in vitro and in vivo bio-imaging based on SERS, fluorescence, photoacoustic and dark-field models. Finally, this article shows perspectives in plasmon-enhanced photothermal and photodynamic therapy.

On paper characterisation of droplet and evaporation study using impedance spectroscopy.

The development of paper-based devices has drawn a significant amount of attention, ranging from the creation of paper electronics to microfluidic devices. The flow of fluids through the paper substrate can be controlled by establishing a variety of barriers, which can be accomplished by either cutting or producing layers that are hydrophobic. Through the utilisation of this feature, a number of investigations, including mixing, modifying, and analytical studies, have been carried out on the paper substrate. However, because of the difficulties associated with its wettability, it is seldom investigated for the purpose of conducting evaporation studies of droplets. Traditionally, evaporation studies are carried out on a solid substrate like glass or silicon. Here we report a paper chip employing an impedance method to determine the characteristics of the droplet. It is also possible to determine the identity of the droplet by utilising the dielectric property of the liquid on a paper chip. A comparison is made between the traditional method of evaporation and the usage of the paper chip for the purpose of studying the evaporation of various liquids, ranging from ionic chemicals to volatile compounds. A subsequent step involves the utilisation of an electrical equivalent circuit in order to acquire the complex system attribute of the evaporation of the cellulose fibres. Finally, this reveals that paper chips have a significant amount of promise for use in scientific applications regarding evaporation analysis.

High sensitivity and automatic chemiluminescence detection of glucose and lactate using a spin-disc paper-based device.

This paper reports a spin-disc paper-based device with 10 individual detection units containing electromagnetic modules controlling the sample incubation time before chemiluminescence (CL) signal detection. After the sample was added to the top paper chip and incubated with the enzyme, the electromagnet was turned off to allow contact between the top and bottom paper. The H2O2 generated by the sample flowed vertically to the bottom paper and initiated the oxidase of the luminol to generate the CL signal. After one detection the disc was automatically rotated to the next position to repeat the above detection. The advantage of using the device over the lateral flow and the in situ detection was firstly proved using the detection of H2O2 and the glucose/lactate sample with 5 minute incubation. The CL intensity was increased 300 times/1000 times as the glucose/lactate was incubated for 5 minutes compared to the non-incubated samples. Afterward, the device was employed to separately detect glucose and lactate diluted in PBS, artificial sweat, artificial saliva, and fresh cell culture media. Finally, the device was employed to detect the glucose and lactate in the media collected over the 24 hour culture of PC3 cells. The uptake and production rates of glucose and lactate were correspondingly determined as 0.328 ± 0.015 pmol h-1 per cell and 1.254 ± 0.053 pmol h-1 per cell, respectively. The reported device has wide application potential due to its capabilities in automatic detection of multiple samples with very high sensitivity and small sample volume (down to 0.5 μL).

Amplification-free detection of Escherichia coli using an acidic deoxyribozyme-based paper device.

We reported a colorimetric paper-based device by integrating the modified acid RNA-cleaving DNAzymes (MaRCD-EC1) for highly sensitive (detection limit = 102 CFU mL-1), and rapid (within 30 min) detection of E. coli without amplification. This device exhibited a clinical sensitivity of 100% and a specificity of 100% in identifying E. coli-associated urinary tract infections (UTIs) using the clinical urine samples.