Air trap and removal on a pressure driven PDMS-based microfluidic device.

With the development of microfluidic technology, microfluidic chips have played a positive role in applications such as cell culture, microfluidic PCR, and nanopore gene sequencing. However, the presence of bubbles interferes with fluid flow and has a significant impact on experimental results. There are many reasons for the generation of bubbles in microfluidic chips, such as pressure changes inside the chip, air vibration inside the chip, and the open chip guiding air into the chip when driving fluid. This study designed and prepared a microfluidic device based on polydimethylsiloxane. First, air was actively introduced into the microfluidic chip, and bubbles were captured through the microfluidic device to simulate the presence of bubbles inside the chip in biological experiments. To remove bubbles trapped in the microfluidic chip, distilled water, distilled water containing surfactants, and mineral oil were pumped into the microfluidic chip. We compared and discussed the bubble removal efficiency under different driving fluids, driving pressures, and open/closed channel configurations. This study helps to understand the mechanism of bubble formation and removal in microfluidic devices, optimize chip structure design and experimental reagent selection, prevent or eliminate bubbles, and reduce the impact of bubbles on experiments.

Rapid-release reversible bonding of PMMA-based microfluidic devices with PBMA coating.

PMMA-based microfluidics have been widely used in various applications in biological and chemical fields. In the fabrication process of PMMA-based microfluidics, the substrate and cover plate usually need to be bonded to enclose the microchannel. The bonding process could be permanent or reversible. In some application scenarios, reversible bonding is needed to retrieve the samples inside the channel or reuse the chip. Current reversible bonding methods for PMMA-based microfluidics usually have drawbacks on bonding strength and contaminations from the adhesives used in the bonding process. In this study, a new approach is proposed for the reversible bonding of PMMA-based microfluidics, a layer of PBMA (with a very similar structure to PMMA) was coated on the surface of PMMA and then use the thermal fusion method to achieve the bonding with a high bonding strength, a tensile bonding strength of around 0.8 MPa was achieved. For debond process, a rapid temperature drop will trigger the immediate release of the bonding within several seconds. Detailed bonding strength measurement and biocompatibility tests were also conducted in this study. The proposed bonding method could have wide application potential in the fabrication of PMMA-based microfluidics.

Integrated sample processing and counting microfluidic device for microplastics analysis.

BACKGROUND: The presence of microplastics is widespread in the ocean, freshwater, soil, or even in the human body. The current microplastics analysis method involves a relatively complicated sieving, digestion filtration, and manual counting process, which is both time-consuming and requires experienced operation personnel. RESULT: This study proposed an integrated microfluidic approach for the quantification of microplastics from river water sediment and biosamples. The proposed two-layer PMMA-based microfluidic device is able to conduct the sample digestion, filtration and counting processes inside the microfluidic chip with the preprogrammed sequence. For demonstration, samples from river water sediment and fish gastrointestinal tract were analyzed, result indicate the proposed microfluidic device is able to perform the quantification of microplastics from river water and biosamples. SIGNIFICANCE AND NOVELTY: Compared with the conventional approach, the proposed microfluidic-based sample processing and quantification method for microplastics are simple, low-cost and with low demand for laboratory equipments, the self-contained system also has the application potential for the continuous on-site inspection of microplastics.

A low-cost and hand-hold PCR microdevice based on water-cooling technology.

Polymerase chain reaction (PCR) has become a powerful tool for detecting various diseases due to its high sensitivity and specificity. However, the long thermocycling time and the bulky system have limited the application of PCR devices in Point-of-care testing. Herein, we have proposed an efficient, low-cost, and hand-hold PCR microdevice, mainly including a control module based on water-cooling technology and an amplification module fabricated by 3D printing. The whole device is tiny and can be easily hand-held with a size of about 110 mm × 100 mm × 40 mm and a weight of about 300 g at a low cost of about $170.83. Based on the water-cooling technology, the device can efficiently perform 30 thermal cycles within 46 min at a heating/cooling rate of 4.0/8.1 ℃/s. To test our instrument, plasmid DNA dilutions were amplified with this device; the results demonstrate successful nucleic acid amplification of the plasmid DNA and exhibit the promise of this device for Point-of-care testing.

Assessment of microplastics using microfluidic approach.

Microplastics are plastic particles smaller than 5 mm, and microplastics have gradually become a severe environmental pollution source that exists in the atmosphere. The identification and quantification of microplastic particles are challenging, current approaches require expensive instruments and are usually time-consuming. In this study, a microfluidic method was introduced to detect and count microplastics using a polymer-based microfluidic chip. Microplastic particles were stained with Nile red, dispersed in the carrier fluid and passed through the microchannel. A fluorescence microscope filmed the whole process as microplastic particles passed through the microchannel. Finally, the software automatically analyzed the video footage for the microplastic particle counting and size analysis. The entire process is fully automated for microplastic particle counting and is much more efficient than the current manual counting method. The proposed study may have broad application potentials in the environmental field.

Fabrication of Transparent and Flexible Digital Microfluidics Devices.

This study proposed a fabrication method for thin, film-based, transparent, and flexible digital microfluidic devices. A series of characterizations were also conducted with the fabricated digital microfluidic devices. For the device fabrication, the electrodes were patterned by laser ablation of 220 nm-thick indium tin oxide (ITO) layer on a 175 µm-thick polyethylene terephthalate (PET) substrate. The electrodes were insulated with a layer of 12 µm-thick polyethylene (PE) film as the dielectric layer, and finally, a surface treatment was conducted on PE film in order to enhance the hydrophobicity. The whole digital microfluidic device has a total thickness of less than 200 µm and is nearly transparent in the visible range. The droplet manipulation with the proposed digital microfluidic device was also achieved. In addition, a series of characterization studies were conducted as follows: the contact angles under different driving voltages, the leakage current density across the patterned electrodes, and the minimum driving voltage with different control algorithms and droplet volume were measured and discussed. The UV-VIS spectrum of the proposed digital microfluidic devices was also provided in order to verify the transparency of the fabricated device. Compared with conventional methods for the fabrication of digital microfluidic devices, which usually have opaque metal/carbon electrodes, the proposed transparent and flexible digital microfluidics could have significant advantages for the observation of the droplets on the digital microfluidic device, especially for colorimetric analysis using the digital microfluidic approach.

CO2 laser fabrication of hydrogel-based open-channel microfluidic devices.

This study proposed a rapid and low-cost fabrication method for open-channel hydrogel-based microfluidic devices using CO2 laser ablation. The agarose hydrogel substrate was prepared with agarose gelation in DI water upon microwave heating, then a commercial CO2 laser system was used for the direct laser ablation of microchannels on the surface of agarose hydrogel substrate, the hydrophilic nature of the microchannels fabricated on hydrogel substrate enables the self-driven of the liquid inside the microchannels with capillary force. The profiles of the laser ablated microchannels on agarose hydrogel substrate with various laser power and scan speed were studied in detail. Due to the loss of water when exposed to the atmosphere, significant deformation of the fabricated microchannels was observed, and the profile change was recorded for 48 h for comparison. An easy-to-access storage method of hydrogel-based microfluidic device in DI water was also proposed in this study. Unlike compact silicon or polymer-based microfluidic devices, the hydrogel is formed by cross-linked polymer chains filled with water, for a better understanding of the diffusion of small molecules into the bulk hydrogel material during fluid propagation inside the microchannel, the Nile red fluorescent was added into the liquid and the diffusion across the hydrogel-based microchannels with time was measured and discussed in this study. Several open-channel agarose hydrogel-based microfluidic devices were fabricated in this study for the demonstration of the proposed fabrication method. The CO2 laser ablation approach for agarose hydrogel-based microfluidic devices has the advantages of rapid processing time, low-cost, highly biocompatible, and self-driven without pumps and could have wide application potentials in biological and medical fields.

Seepage Time Soft Sensor Model of Nonwoven Fabric Based on the Extreme Learning Machine Integrating Monte Carlo.

Nonwoven fiber materials are materials with multifunctional purposes, and are widely used to make masks for preventing the new Coronavirus Disease 2019. Because of the complexity and particularity of their structure, it becomes difficult to model the penetration and flow characteristics of liquid in nonwoven fiber materials. In this paper, a novel seepage time soft sensor model of nonwoven fabric, based on Monte Carlo (MC), integrating extreme learning machine (ELM) (MCELM) is proposed. The Monte Carlo method is used to expand data samples. Then, an ELM method is used to establish the prediction model of the dyeing time of the nonwoven fiber material overlaps with the porous medium, as well as the insertion degree and height of the different quantity of hides. Compared with the back propagation (BP) neural network and radial basis function (RBF) neural network, the results show that the prediction model based on the MCELM method has significant power in terms of accuracy and prediction speed, which is conducive to the precise and rapid manufacture of nonwoven fiber materials in practical applications between liquid seepage characteristics and structural characteristics of porous media. Furthermore, the relationship between the proposed models has certain value for predicting the behavior and use of nonwoven fiber materials with different structural characteristics and related research processes.

Ultra-low-cost fabrication of polymer-based microfluidic devices with diode laser ablation.

In this work, a diode laser ablation approach was used for the fabrication of PMMA-based microfluidic devices. Compared with the conventional CO2 or femtosecond laser fabrication method, the proposed laser ablation method based on diode laser significantly lowered the cost in the fabrication of polymer-based microfluidic devices with comparable resolution and surface quality. PMMA substrate was used for the laser ablation process, due to the transparency of PMMA in the diode laser's working wavelength, a layer of Kraft tape was applied on the surface of PMMA for the absorption of laser energy, and microchannels were then achieved on the surface of PMMA with the proposed low-cost diode laser system. The comparison between the proposed method and the CO2 laser ablation method was also conducted in this study. The profile of the fabricated microchannels was carefully characterized, several microfluidic devices were also fabricated for the demonstration of the proposed fabrication method using a diode laser.

Laser-induced selective wax reflow for paper-based microfluidics.

This study proposes a novel method for the fabrication of paper-based microfluidic devices using laser-induced selective thermal reflow for wax penetration. A layer of wax was evenly deposited on the front side of a filter paper; then a low-cost diode laser was used to scan the designed area from the back side of the filter paper. At the laser irradiated spot, the wax was heated, melted down and penetrated through the whole thickness of the filter paper, and formed hydrophobic barriers on the hydrophilic cellulose fibers. The patterned hydrophobic wax barriers on the filter paper defined the flow path of the fluid for the paper-based microfluidic device. Compared with conventional two-step (deposit and reflow) approaches for paper-based microfluidics using wax barriers, e.g. wax printing, stamping or photolithography, the proposed fabrication protocol achieved wax patterning and reflow simultaneously, conducted during the laser scan process, and without the requirement for any sophisticated instruments or a cleanroom environment. A series of tests were also conducted for the characterization of the proposed paper-based microfluidic device fabrication technique. The fabrication technique used in this approach could have broad application potential in point-of-care diagnosis and testing, especially for applications in the developing world.

Long-term anti-endotoxin/E. coli efficacy in mice transfected with AAV2/1-muBPI25 -muFcγ1.

Bactericidal/permeability increasing (BPI) is an antibiotic protein which kills Gram-negative bacteria and neutralizes endotoxin. We have previously developed a recombinant adeno-associated virus which contains human BPI amino acid residues 1-199 and Fc fragment of human IgG1 gene (AAV-hBPI-Fc) and shown that the recombinant virus can protect mice from lethal endotoxemia. However, whether AAV-hBPI-Fc can be used in vivo for the long term remains unclear. To address this, we established an adeno-associated virus-containing mouse BPI and Fc fragment genes (muBPI-Fc) and compared antigenicity of these recombinant proteins in murine models. Immunohistochemistry showed the expression of both fusion proteins at injected sites. ELISA and Western blotting showed that the muBPI-Fc protein was detected in serum up to 8 weeks after injection, without generation of autoantibodies against muBPI-Fc. In contrast, expressed hBPI-Fc protein was only detected on the 2nd week, whereas the autoantibody against hBPI-Fc protein occurred in serum from the 4th week to the end of study. muBPI-Fc also reduced production of proinflammatory cytokines and protected mice from endotoxemia and bacteremia. Our data showed that AAV-muBPI-Fc has potential long-term efficacy as an anti-endotoxin and has anti-bacterial activity in mice, suggesting the potential clinical application of AAV-hBPI-Fc, such as in endotoxin shock.

T-helper cell type 2 (Th2) memory T cell-potentiating cytokine IL-25 has the potential to promote angiogenesis in asthma.

IL-25 (IL-17E) is a T-helper cell type 2 (Th2) cytokine best described as a potentiator of Th2 memory responses. Reports of expression of its receptor, IL-25R, on airways structural cells suggest a wider role for IL-25 in remodeling. We hypothesized that IL-25 stimulates local angiogenesis in the asthmatic bronchial mucosa. Immunoreactive IL-25(+), IL-25R(+), and CD31(+) (endothelial) cells in sections of bronchial biopsies from asthmatics and controls were detected by immunohistochemistry. The effect of IL-25 on angiogenesis was examined using an in vitro assay. Real-time PCR was used to detect expression of IL-25R and VEGF mRNA in cultured human vascular endothelial cells (HUVEC), and a cell proliferation kit (WST-8) was used to measure the effect of IL-25 on HUVEC proliferation. Immunostaining showed that IL-25(+), IL-25R(+), and CD31(+)/IL-25R(+) cells were significantly elevated in the bronchial mucosa of asthmatics compared with controls (P < 0.003). In asthmatics, the numbers of IL-25(+) cells correlated inversely with the forced expiratory volume in 1 s (r = -0.639; P = 0.01). In vitro, HUVEC constitutively expressed IL-25R, which was up-regulated further by TNF-α. IL-25 and TNF-α also increased expression of VEGF and VEGF receptors. IL-25 increased HUVEC proliferation and the number, length, and area of microvessel structures in a concentration-dependent manner in vitro. VEGF blockade, the PI3K-specific inhibitor LY294002, and the MAPK/ERK1/2 (MEK1/2)-specific inhibitor U0126 all markedly attenuated IL-25-induced angiogenesis, and the inhibitors also reduced IL-25-induced proliferation and VEGF expression. Our findings suggest that IL-25 is elevated in asthma and contributes to angiogenesis, at least partly by increasing endothelial cell VEGF/VEGF receptor expression through PI3K/Akt and Erk/MAPK pathways.