Microfluidic Analytical Devices Research Articles

Microfluidic Thread-Based Analytical Platform for Reaction Screening Enabled by In Situ Thread Spray Mass Spectrometry.

This study describes a microfluidic thread-based analytical device (μTAD) capable of in situ mass spectrometric analysis for continuous flow reaction monitoring. Organic reaction screening is foundational to drug discovery. Microfluidic devices are of special interest here because they provide continuous reaction monitoring with advantages such as the use of smaller reagent volumes and short analysis times. We created a μTAD platform from cellulose thread and a microscope glass slide in a Y-shaped format, which enabled multiple reactants to be introduced through individual channels/reservoirs. The naturally corrugated and intertwined subfibers within cellulose thread substrates serve as serpentine mixing medium, requiring no special preparation. The capillary action behind the flow of the reactants in the thread substrate leads to controlled solvent evaporation, which is found to accelerate the chemical reactions. When performed in front of a mass spectrometer, the μTAD platform provides a facile approach to directly characterize the reaction products formed on the thread substrate. We have used the μTAD mass spectrometry platform to monitor (1) Katritzky chemistry for the synthesis of pyridinium species, (2) aza-Michael addition for uncatalyzed formation of C-N bonds, and (3) Mannich reaction, which is a three-component condensation reaction that affords β-amino carbonyl compounds. Similar reactions were screened using nanoelectrospray ionization, which suggested limited reactivity in charged microdroplets. Therefore, the main factors observed to influence reactivity on the μTAD platform were identified to include effective mixing and solvent evaporation, leading to reagent concentration in the resultant thin films. The μTAD proved to be a rapid, efficient, and low-cost platform for organic reaction monitoring, which has the potential to act as an alternative to the traditional batch screening method.

Fe-Doped Red Fluorescent Carbon Dots for Caffeine Analysis in Energy Drinks Using a Paper-Based Sensor.

This study introduces a highly sensitive and selective method for detecting caffeine in energy drinks by using red florescence iron and nitrogen co doped carbon dots (Fe-NCDs) as a florescent prob. The Fe-NCDs were synthesized by using an eco-friendly hydrothermal. Providing uniform, quasi-spherical nanoparticles. The photoluminescence properties of the Fe-NCDs exhibit strong red emission making them suitable for fluorescence-based sensing. A microfluidic paper analytical device (µPAD) was developed and coupled with a smartphone-based detection system to facilitate portable, low-cost caffeine quantification. The Fe-NCDs were embedded in the µPADs, enabling fluorescence enhancement upon interaction with caffeine. This enhancement was quantitatively analyzed using the smartphone camera and ImageJ software, revealing a strong linear correlation in the range of 1 to 40µg/mL when both Gray Value (G.V) and Red-Green-Blue (RGB) of reaction analyzed by the software. The limit of detection (LOD) was of 0.024µg/mL and 0.032µg/mL respectively for both applied principles. The methods indicate remarkable selectivity for caffeine, and was validated through accurate recovery studies in commercial samples. This innovative method provides a powerful, cost-effective, and environmentally sustainable solution for on-site caffeine detection in energy drinks, offering significant potential for application in food safety and quality control.

A Spectroscopy Solution for Contactless Conductivity Detection in Capillary Electrophoresis.

This paper introduces a novel contactless single-chip detector that utilizes impedance-to-digital conversion technology to measure impedance in the microfluidic channel or capillary format analytical device. The detector is designed to operate similarly to capacitively coupled contactless conductivity detectors for capillary electrophoresis or chromatography but with the added capability of performing frequency sweeps up to 200 kHz. At each recorded data point, impedance and phase-shift data can be extracted, which can be used to generate impedance versus frequency plots, or phase-shift versus frequency plots. Real and imaginary parts can also be calculated from the data, allowing for the generation of Nyquist diagrams. This detector represents the first of its kind in the contactless conductivity class to provide spectrum-type data, as demonstrated in capillary electrophoresis experiments.

Development of paper-based microfluidic analytical device (μPAD) for the determination of paracetamol in water samples: Optimization using response surface methodology (RSM)

Detecting and quantifying pharmaceutical compounds in various environmental matrices is complex and challenging. This difficulty stems from the trace levels at which these compounds are found and the lack of analytical methods that are rapid, cost-effective, and portable. To address these challenges, this study aimed to develop microfluidic paper-based analytical devices (μ-PADs) using beeswax screen printing for fabrication. Key parameters, including reaction time, concentration, reagent volume, and channel length, were optimized using response surface methodology. Under optimal conditions of 5 ppm sample concentration, 10 μL reagent volume, 10 min reaction time, and 2 cm channel length, the analytical performance of the μPAD was evaluated and compared with the standard UV–Vis spectrophotometry method. The microfluidic analytical device demonstrated detection limits at 0.03 μg/ml, compared to 0.01 μg/ml for the UV–Vis spectrophotometer. Although the sensitivity of µ-PADs in this study (0.03 μg/ml) is lower than that of UV–Vis (0.01 μg/ml), it represents an improvement over the previous µ-PAD report (1 μg/ml) on the same analytes. Both methods exhibited commendable precision, with a relative standard deviation below 2%. Additionally, recovery rates were acceptable and comparable, ranging from 86.8 to 99.6% for µ-PADs and 96.5–99% for UV–Vis. The analytical performance evaluation suggests that µPADs provide excellent sensitivity, precision, and accuracy for trace-level paracetamol analysis. A paired t-test further confirmed no statistically significant difference between the two methods, underscoring the promising potential of µ-PADs for trace-level paracetamol quantification in water samples without conventional analytical instruments.

Adhered-3D paper microfluidic analytical device based on oxidase-mimicking activity of Co-doped carbon dots nanozyme for point-of-care testing of alkaline phosphatase

Adhered-3D paper microfluidic analytical device based on oxidase-mimicking activity of Co-doped carbon dots nanozyme for point-of-care testing of alkaline phosphatase

Advances in textile-based microfluidics for biomolecule sensing.

Textile-based microfluidic biosensors represent an innovative fusion of various multidisciplinary fields, including bioelectronics, material sciences, and microfluidics. Their potential in biomedicine is significant as they leverage textiles to achieve high demands of biocompatibility with the human body and conform to the irregular surfaces of the body. In the field of microfluidics, fabric coated with hydrophobic materials serves as channels through which liquids are transferred in precise amounts to the sensing element, which in this case is a biosensor. This paper presents a condensed overview of the current developments in textile-based microfluidics and biosensors in biomedical applications over the past 20 years (2005-2024). A literature search was performed using the Scopus database. The fabrication techniques and materials used are discussed in this paper, as these will be key in various modifications and advancements in textile-based microfluidics. Furthermore, we also address the gaps in the application of textile-based microfluidic analytical devices in biomedicine and discuss the potential solutions. Advances in textile-based microfluidics are enabled by various printing and fabric manufacturing techniques, such as screen printing, embroidery, and weaving. Integration of these devices into everyday clothing holds promise for future vital sign monitoring, such as glucose, albumin, lactate, and ion levels, as well as early detection of hereditary diseases through gene detection. Although most testing currently takes place in a laboratory or controlled environment, this field is rapidly evolving and pushing the boundaries of biomedicine, improving the quality of human life.

Automatic characterization of capillary flow profile of liquid samples on μTADs based on capacitance measurement

Capillary flow profile of liquid samples in porous media is closely related to the important properties of liquid samples, including the viscosity and the surface energy. Therefore, capillary flow profile can be used as an index to differentiate liquid samples with different properties. Fast and automatic characterization of capillary flow profile of liquid samples is necessary. In this work, we develop a portable and economical capacitance acquisition system (CASY) to easily obtain the capillary flow profile of liquid samples on microfluidic thread-based analytical devices (μTADs) by measuring the capacitance during the capillary flow. At first, we validate the accuracy of this method by comparing with the traditional method by video analysis in obtaining the capillary flow profiles in μTADs of cotton threads or glass fiber threads. Then we use it to differentiate liquid samples with different viscosity (mixture of water and glycerol). In addition, capillary flow profile on μTADs with chemical valves (chitosan or sucrose) can also be obtained on this device. Lastly, we show the potential of this device in measurement of hematocrit (HCT) of whole blood samples. This device can be used to catalog liquid biological samples with different properties in point-of-care diagnostics in the near future.

Paper chip-based colorimetric biosensor for glucose by using CuFe2O4@GO as nanozyme

Paper chip-based colorimetric biosensor for glucose by using CuFe2O4@GO as nanozyme

Copper (II)-catalyzed polydopamine mediated photothermal sensors for visual quantitative point-of-care testing

Temperature sensing is commonly used in point-of-care (POC) detection technologies, yet the portability and convenience of use are frequently compromised by the complexity of thermosensitive processes and signal transduction. Especially, multi-step target recognition reactions and temperature measurement in the reaction vessel present challenges in terms of stability and integration of detection devices. To further combine photothermal reaction and signal readout in one assay, these two processes enable to be integrated into miniaturized microfluidic chips, thereby facilitating photothermal sensing and achieving a simple visual temperature sensing as POC detection. A copper ion (Cu2+)-catalyzed photothermal sensing system integrated onto a microfluidic distance-based analytical device (μDAD), enabling the visual, portable, and sensitive quantitative detection of multiple targets, including ascorbic acid, glutathione, and alkaline phosphatase (ALP). The polydopamine nanoparticles (PDA NPs) were synthesized by the regulation of free Cu2+ through redox or coordination reactions, facilitating the transduction of distinct photothermal response signals and providing the versatile Cu2+-responsive sensing systems. Promoted by integration with a photothermal μDAD, the system combines PDA's photothermal responsiveness and thermosensitive gas production of ammonium bicarbonate for improved sensitivity of ALP detection, reaching the detection limit of 9.1 mU/L. The system has successfully achieved on-chip detection of ALP with superior anti-interference capability and recoveries ranging from 96.8% to 104.7%, alongside relative standard deviations below 8.0%. The μDAD design accommodated both the photothermal reaction of PDA NPs and thermosensitive gas production reaction, achieving the rapid sensing of visual distance signals. The μDAD-based Cu2+-catalyzed photothermal sensing system holds substantial potential for applications in biochemical analysis and clinical diagnostics, underscored by the versatile Cu2+ regulation mechanism for a broad spectrum of biomarkers.

Paper-Based Microfluidic Analytical Device Patterned by Label Printer for Point-of-Care Blood Glucose and Hematocrit Detection Using 3D-Printed Smartphone Cassette.

This study presents a portable, low-cost, point-of-care (POC) system for the simultaneous detection of blood glucose and hematocrit. The system consists of a disposable origami microfluidic paper-based analytical device (μPAD) for plasma separation, filtration, and reaction functions and a 3D-printed cassette for hematocrit and blood glucose detection using a smartphone. The origami μPAD is patterned using a cost-effective label printing technique instead of the conventional wax printing method. The 3D-printed cassette incorporates an array of LED lights, which mitigates the effects of intensity variations in the ambient light and hence improves the accuracy of the blood glucose and hematocrit concentration measurements. The hematocrit concentration is determined quantitatively by measuring the distance of plasma wicking along the upper layer of the origami μPAD, which is pretreated with sodium chloride and Tween 20 to induce dehydration and aggregation of the red blood cells. The filtered plasma also penetrates to the lower layer of the origami μPAD, where it reacts with embedded colorimetric assay reagents to produce a yellowish-brown complex. A color image of the reaction complex is captured using a smartphone inserted into the 3D-printed cassette. The image is analyzed using self-written RGB software to quantify the blood glucose concentration. The calibration results indicate that the proposed detection platform provides an accurate assessment of the blood glucose level over the range of 45-630 mg/dL (R2 = 0.9958). The practical feasibility of the proposed platform is demonstrated by measuring the blood glucose and hematocrit concentrations in 13 human whole blood samples. Taking the measurements obtained from commercial glucose and hematocrit meters as a benchmark, the proposed system has a differential of no more than 6.4% for blood glucose detection and 9.1% for hematocrit detection. Overall, the results confirm that the proposed μPAD is a promising solution for cost-effective and reliable POC health monitoring.

Multifunctional Metal-Organic Frameworks Driven Three-Dimensional Folded Paper-Based Microfluidic Analysis Device for Chlorpyrifos Detection.

Chlorpyrifos (CPF) residues in food pose a serious threat to ecosystems and human health. Herein, we propose a three-dimensional folded paper-based microfluidic analysis device (3D-μPAD) based on multifunctional metal-organic frameworks, which can achieve rapid quantitative detection of CPF by fluorescence-colorimetric dual-mode readout. Upconversion nanomaterials were first coupled with a bimetal organic framework possessing peroxidase activity to create a fluorescence-quenched nanoprobe. After that, the 3D-μPAD was finished by loading the nanoprobe onto the paper-based detection zone and spraying it with a color-developing solution. With CPF present, the fluorescence intensity of the detection zone gradually recovers, the color changes from colorless to blue. This showed a good linear relationship with the concentration of CPF, and the limits of detection were 0.028 (fluorescence) and 0.043 (colorimetric) ng/mL, respectively. Moreover, the 3D-μPAD was well applied in detecting real samples with no significant difference compared with the high-performance liquid chromatography method. We believe it has huge potential for application in the on-site detection of food hazardous substance residues.

Microfluidic paper analytic device (μPAD) technology for food safety applications.

Foodborne pathogens, food adulterants, allergens, and toxic chemicals in food can cause major health hazards to humans and animals. Stringent quality control measures at all stages of food processing are required to ensure food safety. There is, therefore, a global need for affordable, reliable, and rapid tests that can be conducted at different process steps and processing sites, spanning the range from the sourcing of food to the end-product acquired by the consumer. Current laboratory-based food quality control tests are well established, but many are not suitable for rapid on-site investigations and are costly. Microfluidic paper analytical devices (μPADs) are a fast-growing field in medical diagnostics that can fill these gaps. In this review, we describe the latest developments in the applications of microfluidic paper analytic device (μPAD) technology in the food safety sector. State-of-the-art μPAD designs and fabrication methods, microfluidic assay principles, and various types of μPAD devices with food-specific applications are discussed. We have identified the prominent research and development trends and future directions for maximizing the value of microfluidic technology in the food sector and have highlighted key areas for improvement. We conclude that the μPAD technology is promising in food safety applications by using novel materials and improved methods to enhance the sensitivity and specificity of the assays, with low cost.

A miniaturized additive-manufactured carbon black/PLA electrochemical sensor for pharmaceuticals detection

Additive manufacturing is a technique that allows the construction of prototypes and has evolved a lot in the last 20 years, innovating industrial fabrication processes in several areas. In chemistry, additive manufacturing has been used in several functionalities, such as microfluidic analytical devices, energy storage devices, and electrochemical sensors. Theophylline and paracetamol are important pharmaceutical drugs where overdosing can cause adverse effects, such as tachycardia, seizures, and even renal failure. Therefore, this paper aims at the development of miniaturized electrochemical sensors using 3D printing and polylactic acid-based conductive carbon black commercial filament for theophylline and paracetamol detection. Electrochemical characterizations of the proposed sensor were performed to prove the functionality of the device. Morphological characterizations were carried out, in which chemical treatment could change the surface structure, causing the improvement of the analytical signal. Thus, the detection of theophylline at a linear range of 5.00-150μmolL-1 with a limit of detection of 1.2μmolL-1 was attained, and the detection of paracetamol at a linear range of 1.00-200μmolL-1 with a limit of detection of 0.370μmolL-1 was obtained, demonstrating the proposed sensor effectively detected pharmaceutical drugs.

Construction of Out-of-Equilibrium Metabolic Networks in Nano- and Micrometer-Sized Vesicles.

We present a method to incorporate into vesicles complex protein networks, involving integral membrane proteins, enzymes, and fluorescence-based sensors, using purified components. This method is relevant for the design and construction of bioreactors and the study of complex out-of-equilibrium metabolic reaction networks. We start by reconstituting (multiple) membrane proteins into large unilamellar vesicles (LUVs) according to a previously developed protocol. We then encapsulate a mixture of purified enzymes, metabolites, and fluorescence-based sensors (fluorescent proteins or dyes) via freeze-thaw-extrusion and remove non-incorporated components by centrifugation and/or size-exclusion chromatography. The performance of the metabolic networks is measured in real time by monitoring the ATP/ADP ratio, metabolite concentration, internal pH, or other parameters by fluorescence readout. Our membrane protein-containing vesicles of 100-400 nm diameter can be converted into giant-unilamellar vesicles (GUVs), using existing but optimized procedures. The approach enables the inclusion of soluble components (enzymes, metabolites, sensors) into micrometer-size vesicles, thus upscaling the volume of the bioreactors by orders of magnitude. The metabolic network containing GUVs aretrapped in microfluidic devices for analysis by optical microscopy.

Microfluidic-oriented green synthesis of pepsin-doped gold nanoparticles for colorimetric and photothermal dual-readout detection of food hazards

Gold nanoparticles (AuNPs)-based immunochromatographic assay has gained popularity as a rapid detection method for food hazards. Synthesizing highly stable AuNPs in a rapid, simple and environmentally friendly manner is a key focus in this field. Here, we present a green microfluidic strategy for the rapid, automated, and size-controllable synthesis of pepsin-doped AuNPs (AuNPs@Pep) by employing glucose-pepsin as a versatile reducing agent and stabilizer. Through combining the colorimetric and photothermal (PoT) properties of AuNPs@Pep, both "signal-off" and "signal-on" formats of microfluidic paper analytical devices (PADs) were developed for detection of a small molecule antibiotic, florfenicol, and an egg allergen, ovalbumin. Compared to the colorimetric mode, a 4-fold and 3-fold improvement in limit of detection was observed in the "signal-off" detection of florfenicol and the "signal-on" detection of ovalbumin, respectively. The results demonstrated the practicality of AuNPs@Pep as a colorimetric/PoT dual-readout probe for immunochromatographic detection of food hazards at different molecular scales.

Microfluidic Immunosensing Platform Based on a Rolling Circle Amplification-Assisted DNA Dendrimer Probe for Portable and Sensitive Detection of Allergen-Specific IgE.

The robust point-of-care platform for sensitive, multiplexed, and affordable detection of allergen-specific IgE (sIgE) is an urgent demand in component-resolved diagnostics. Here, we developed a microfluidic immunosensing platform based on a rolling circle amplification-assisted DNA dendrimer probe for sensitive detection of multiple sIgEs. The versatile multichannel microfluidic whole blood analytical device integrates cell filtration, recombinant antigen-modified magnetic enrichment, and DNA dendrimer probe-amplified signal transduction for portable on-chip analysis. Three sIgEs against common oyster allergens were simultaneously detected in blood samples by simple smartphone-based imaging without any pretreatment. The quantitative detection of multiple allergen-specific antibodies on the platform was achieved with limits of detection of less than 50 pg/mL, exhibiting superior sensitivity compared to most point-of-care testing. The detection results of 55 serum samples and 4 whole blood samples were 100% consistent with the ELISA results, confirming the accuracy and stability of our platform. Additionally, the reversible combination of hexahistidine6-tag and Ni-IMAC magbead was elegantly utilized on the immunosensing platform for desired reversibility. With the advantages of general applicability, high sensitivity, and reversibility, the DNA dendrimer-based microfluidic immunosensing platform provides great potential for the portable detection of immune proteins as a point-of-care platform in disease diagnostics and biological analysis.

Cotton threads encapsulated by thermal contraction tube for point-of-care diagnostics

Cotton threads encapsulated by thermal contraction tube for point-of-care diagnostics

Development of a 3D Microfluidic Analytical Device for the Detection of Pathogenic Bacteria in Commercial Food Samples with Loop-Mediated Isothermal Amplification

Traditional methods of detecting foodborne pathogens take several days to produce the required results. Furthermore, various molecular techniques (e.g., PCR) that also produce reliable results in the detection of pathogenic bacteria have been introduced, but the cost–time ratio required does not allow them to be considered a substantial solution to this specific problem. Three-dimensional (3D) printing technology provides the ability to design and manufacture microfluidic analytical devices using conventional 3D printers, which, in combination with colorimetric loop-mediated isothermal amplification (LAMP), may further simplify the process. The overall reduction in time and cost may provide the opportunity to upscale this diagnostic modality. Moreover, unlike most microfluidic analytical devices, this technique is simpler and more user-friendly, as it does not require any expertise or additional equipment apart from a conventional oven. A 3D-printed microfluidic analytical device in combination with LAMP was developed and tested for the simultaneous detection of foodborne pathogens in food samples. A total of 150 commercial food specimens (50 milk, 50 chicken, 50 lettuce samples) were analyzed for possible contamination with Salmonella typhimurium, Listeria monocytogenes and Escherichia coli. The 3D-printed microfluidic device was 100% precise for both negative (80 samples) and positive samples (7 samples were positive for S. typhimurium, 28 for L. monocytogenes, and 35 for E. coli) for all pathogens. Overall, the amount of data analyzed led to a high level of confidence in the precision of this device. As such, this new 3D device in combination with LAMP provides a precise detection method for food pathogens with a low detection limit.

NFC-enabled photothermal-based microfluidic paper analytical device for glucose detection.

This study introduces the development of a photothermal-based microfluidic paper analytical device (PT-µPAD) integrated with near-field communication (NFC) technology and smartphone readout for enzyme-free glucose quantification in human samples. With the properties of gold nanoparticles (AuNPs) both as a nanozyme and as a photothermal substrate, there is no need for costly reagents like enzymes or a readout instrumentation for the selective and sensitive detection of glucose. In PT-µPADs, AuNPs are etched by hydrogen peroxide (H2O2) generated from glucose catalysis. Photothermal detection from the plasmonic heating of these AuNPs when illuminated by a 533nm LED light source is achieved by inserting the PT-µPAD sensor into a portable NFC platform suitable for smartphone readout. Temperature variation is directly proportional to the glucose concentration. After optimization, we acquired a linear range between 5.0 and 20.0 µmol L-1 (R2 = 0.9967) and a limit of detection (LOD) of 25.0 nmol L-1 for glucose. Additionally, while our sensor does not utilize any enzyme, it is remarkably selective to glucose with no effects from interferences. Recovery studies in various human control samples indicated a range of 99.73-102.66% with the highest RSD of 3.53%, making it highly accurate and precise. Moreover, our method is more sensitive than other methods relying on conventional µPADs for glucose sensing. By integrating the potential benefits of microfluidics, nanomaterials as nanozymes, and NFC technology for wireless readout, our sensor demonstrates great promise as an accessible, affordable, and shelf-stable device for glucose quantification. Moreover, this concept can be extended to detect other molecules of interest as a point-of-care (POC) diagnostics device.

Humidity-enhanced microfluidic plasma separation on Chinese Xuan-papers.

The first step in blood testing necessitates blood separation to obtain an adequate volume of plasma. Traditional centrifugation is bulky, expensive and electricity-powered, which is not suitable for micro-scale blood plasma separation in point-of-care testing (POCT) cases. Microfluidic paper-based plasma separation devices present a promising alternative for plasma separation in such occasions. However, they are limited in terms of plasma yield, which hinders analyte detection. Herein, we proposed a humidity-enhanced paper-based microfluidic plasma separation method to address this issue. Specifically, paper was first treated by blood-typing antibodies, then samples of whole blood were introduced into the prepared paper. After waiting for 5 min for RBC agglutination and plasma wicking under high humidity, micro-scale plasma separation from whole blood was achieved. As a result, an extremely high plasma yield of up to 60.1% could be separated from whole blood through using Xuan-paper. Meanwhile, the purity of plasma could reach 99.99%. Finally, this innovative approach was effortlessly integrated into distance-based glucose concentration detection, enabling rapid determination of blood glucose levels through naked-eye observation. Considering the simplicity and inexpensiveness of this method, we believe that this technology could be integrated to more paper-based microfluidic analytical devices for rapid and accurate detection of plasma analytes in POCT.