Ellen Flávia Moreira Gabriel

Rational selection of substrates to improve color intensity and uniformity on microfluidic paper-based analytical devices

A systematic investigation was conducted to study the effect of paper type on the analytical performance of a series of microfluidic paper-based analytical devices (μPADs) fabricated using a CO2 laser engraver. Samples included three different grades of Whatman chromatography paper, and three grades of Whatman filter paper. According to the data collected and the characterization performed, different papers offer a wide range of flow rate, thickness, and pore size. After optimizing the channel widths on the μPAD, the focus of this study was directed towards the color intensity and color uniformity formed during a colorimetric enzymatic reaction. According to the results herein described, the type of paper and the volume of reagents dispensed in each detection zone can determine the color intensity and uniformity. Therefore, the objective of this communication is to provide rational guidelines for the selection of paper substrates for the fabrication of μPADs.

Modification of microfluidic paper-based devices with silica nanoparticles

This paper describes a silica nanoparticle-modified microfluidic paper-based analytical device (μPAD) with improved color intensity and uniformity for three different enzymatic reactions with clinical relevance (lactate, glucose, and glutamate). The μPADs were produced on a Whatman grade 1 filter paper and using a CO2 laser engraver. Silica nanoparticles modified with 3-aminopropyltriethoxysilane were then added to the paper devices to facilitate the adsorption of selected enzymes and prevent the washing away effect that creates color gradients in the colorimetric measurements. According to the results herein described, the addition of silica nanoparticles yielded significant improvements in color intensity and uniformity. The resulting μPADs allowed for the detection of the three analytes in clinically relevant concentration ranges with limits of detection (LODs) of 0.63 mM, 0.50 mM, and 0.25 mM for lactate, glucose, and glutamate, respectively. An example of an analytical application has been demonstrated for the semi-quantitative detection of all three analytes in artificial urine. The results demonstrate the potential of silica nanoparticles to avoid the washing away effect and improve the color uniformity and intensity in colorimetric bioassays performed on μPADs.

Fast and versatile fabrication of PMMA microchip electrophoretic devices by laser engraving: CE and CEC

This paper describes the effects of different modes and engraving parameters on the dimensions of microfluidic structures produced in PMMA using laser engraving. The engraving modes included raster and vector, while the explored engraving parameters included power, speed, frequency, resolution, line‐width, and number of passes. Under the optimum conditions, the technique was applied to produce channels suitable for CE separations. Taking advantage of the possibility to cut‐through the substrates, the laser was also used to define solution reservoirs (buffer, sample, and waste) and a PDMS‐based decoupler. The final device was used to perform the analysis of a model mixture of phenolic compounds within 200 s with baseline resolution.

Polyester‐toner electrophoresis microchips with improved analytical performance and extended lifetime

This paper reports the fabrication of polyester‐toner (PT) electrophoresis microchips with improved analytical performance and extended lifetime. This has been achieved with a better understanding about the EOF generation and the influence of some parameters including the channel dimensions (width and depth), the injection mode, and the addition of organic solvent to the running buffer. The analytical performance of the PT devices was investigated using a capacitively coupled contactless conductivity detector and inorganic cations as model analytes. The proposed devices have exhibited EOF values of (3.4 ± 0.2) × 10−4 cm2 V−1 s−1 with good stability over 25 consecutive runs. It has been found that the EOF magnitude depends on the channel dimension, i.e. the wider the channel, the higher the EOF value. The separation efficiency for inorganic cations ranged from 13 000 to 50 000 plates/m. The LOD found for K+, Na+, and Li+ were 4.2, 7.3, and 23 μM, respectively. In addition, the same PT device has been used by three consecutive days. Lately, due to improved analytical performance, it was carried out by the first time the detection of inorganic cations in real samples such as energetic drinks and pharmaceutical formulations.

Paper-Based Colorimetric Biosensor for Tear Glucose Measurements

This paper describes a paper-based colorimetric biosensor for measuring glucose concentration levels in human tear samples. Colorimetric biosensors were wax printed on paper platforms and modified with chitosan previously prepared in acetic acid. The proposed device was explored to measure the glucose levels in human tear samples using 3,3′,5,5′-tetramethylbenzydine (TMB) as the chromogenic reagent. The paper-based colorimetric biosensor exhibited a linear behavior for the glucose concentration range between 0.1 and 1.0 mM. The achieved analytical sensitivity and limit of detection (LOD) were 84 AU/mM and 50 µM, respectively. Moreover, the device provided analytical reliability and no statistical difference when compared to the data recorded with a commercial glucometer. The proof-of-concept of our device was successfully demonstrated by measuring the glucose levels in six tear samples from nondiabetic subjects. In general, the results showed that the colorimetric biosensor has noticeable potential to be used as a powerful tool for tear glucose monitoring, since this fluid offers lower potential interferences, non-invasive sample collection and is pain-free. Furthermore, the proposed device could facilitate the treatment of diabetic patients who need constant control of glucose levels and cannot tolerate multiple finger sticks per day.

Characterization of microchip electrophoresis devices fabricated by direct-printing process with colored toner

This paper reports for the first time the use of colored toner to produce polyester toner (PT) ME devices. Colored PT devices were designed in drawing software and printed on a polyester film using a color laser printer with 3600 dpi resolution. The colored toner is composed of a copolymer mixture (styrene and acrylate), wax, silicon dioxide, and pigments. The presence of silica in the toner composition has enhanced the EOF magnitude and improved the analytical performance. For a pH range between 2 and 12, the EOF measured on a magenta PT chip, for example, ranged from 3.8 to 5.8 (× 10−4 cm2 V−1 s−1). Typical separations of inorganic cations (K+, Na+, and Li+) were used as model system to investigate the analytical feasibility of the proposed devices. The repeatability for the migration times of all analytes exhibited RSD values lower than 1% (n = 10). The separation efficiencies found on colored PT devices ranged from 10 000 to 49 000 plates/m, which means between 7 and 23% of the maximum theoretical efficiency on this microfluidic platform (1.85 × 105 plates/m). The improvements achieved on the proposed devices are associated with the small additional amount of silica on the toner composition as well as the printing of channels with smoother surfaces and better uniformity when compared to the conventional PT chips printed with monochromatic laser printers.

Simple, rapid and, cost‐effective fabrication of PDMS electrophoresis microchips using poly(vinyl acetate) as photoresist master

This study describes a simple, rapid, and cost‐effective fabrication of PDMS electrophoresis microchips using poly(vinyl acetate) (PVAc) emulsion as photoresist master. High‐relief microfluidic structures were defined on poly(vinyl acetate) previously deposited on printed circuit boards surfaces without cleanroom facilities and sophisticated instrumentation. After a UV exposure, channels with heights ranging from 30 to 140 μm were obtained by controlling the emulsion mass deposited on the master surface. The developing stage was performed using water rather than the organic solvents that are applied for conventional masks. The surface morphology was characterized by optical imaging, profilometry, and SEM. Based on the achieved results, the proposed method offers suitable reproducibility for the prototyping of electrophoresis microchips in PDMS. The feasibility of the resulting PDMS electrophoresis chips was successfully demonstrated with the separation of major inorganic cations within 100 s using a contactless conductivity detection system. The separation efficiencies ranged from ca. 67 900 to 125 600 plates/m. Due to the satisfactory performance and simplified instrumentation, we believe this fabrication protocol presents potential to be implemented in any chemical, biochemical, or biological laboratory.

Hydrodynamic injection on electrophoresis microchips using an electronic micropipette

Here we report for the first time the use of an electronic micropipette as hydrodynamic (HD) injector for microchip electrophoresis (ME) devices. The micropipette was directly coupled to a PDMS device, which had been fabricated in a simple cross format with two auxiliary channels for sample volume splitting. Sample flow during the injection procedure was controlled in automatic dispenser mode using a volume of 0.6µL. Channel width and device configuration were optimized and the best results were achieved using a simple cross layout containing two auxiliary channels with 300µm width for sample splitting. The performance of the HD injector was evaluated using a model mixture of high-mobility cationic species. The results obtained were compared to the data obtained via electrokinetic (EK) injection. Overall, the HD provided better analytical performance in terms of resolution and injection-to-injection repeatability. The relative standard deviation (RSD) values for peak intensities were lower than 5% (n=10) when the micropipette was employed. In comparison with EK injection, the use of the proposed HD injector revealed an unbiased profile for a mixture containing K+ and Li+(300 µmol L-1 each) over various buffer concentrations. For EK injection, the peak areas decreased from 2.92 ± 0.20-0.72 ± 0.14Vs for K+ and from 1.30 ± 0.10-0.38 ± 0.10Vs for Li+ when the running buffer increased from 20 to 50mmolL-1. For HD injection, the peak areas for K+ and Li+ exhibited average values of 2.48±0.07 and 2.10±0.06Vs, respectively. The limits of detection (LDs) for K+, Na+ and Li+ ranged from 18 to 23µmolL-1. HD injection through an electronic micropipette allows to automatically dispense a bias-free amount of sample inside microchannels with acceptable repeatability. The proposed approach also exhibited instrumental simplicity, portability and minimal microfabrication requirements.

Recent advances in toner-based microfluidic devices for bioanalytical applications

Toner-based microfluidic devices have emerged since 2003 as promising platforms for bioanalytical applications. The simplicity of the fabrication process, the low instrumental requirements and the global affordability of required consumables are some key features that encourage the use of this kind of substrate for chemical and biochemical assays. This review aims to cover the recent advances regarding the fabrication procedures involving the laser printing of microfluidic devices on polyester films as well as the association of printing, laser cutting and lamination to create devices assembled in a multi-layer architecture. Examples of applications involving electrophoresis, mixing, concentration, purification, extraction, amplification, rotation-driven fluidic transport, enzyme-linked immunoassays and colorimetric lateral assays on toner-based platforms will be presented and discussed. Lastly, the versatility of the toner-based fabrication technique to enable the production of hydrophobic valves, droplet generator devices, and mimetic vein-on-a-chip and hybrid devices will also be covered.

Enhanced Performance of Colorimetric Biosensing on Paper Microfluidic Platforms Through Chemical Modification and Incorporation of Nanoparticles

This chapter describes two different methodologies used to improve the analytical performance of colorimetric paper-based biosensors. Microfluidic paper-based analytical devices (μPADs) have been produced by a stamping process and CO2 laser ablation and modified, respectively, through an oxidation step and incorporation of silica nanoparticles on the paper structure. Both methods are employed in order to overcome the largest problem associated with colorimetric detection, the heterogeneity of the color distribution in the detection zones. The modification steps are necessary to improve the interaction between the paper surface and the selected enzymes. The enhanced performance has ensured reliability for quantitative analysis of clinically relevant compounds.