Advances in Microfluidics and Lab on a Chip Technology

The eutrophication of aquatic ecosystems caused by rapid human urbanization has led to an increased production of potentially hazardous bacterial populations, known as blooms. One of the most notorious forms of these aquatic blooms are cyanobacteria, which in sufficiently large quantities can pose a hazard to human health through ingestion or prolonged exposure. Currently, one of the greatest difficulties in regulating and monitoring these potential hazards is the early detection of cyanobacterial blooms, in real time. Therefore, this paper presents an integrated microflow cytometry platform for label-free phycocyanin fluorescence detection, which can be used for the rapid quantification of low-level cyanobacteria and provide early warning alerts for potential harmful cyanobacterial blooms. An automated cyanobacterial concentration and recovery system (ACCRS) was developed and optimized to reduce the assay volume, from 1000 mL to 1 mL, to act as a pre-concentrator and subsequently enhance the detection limit. The microflow cytometry platform utilizes an on-chip laser-facilitated detection to measure the in vivo fluorescence emitted from each individual cyanobacterial cell, as opposed to measuring overall fluorescence of the whole sample, potentially decreasing the detection limit. By applying transit time and amplitude thresholds, the proposed cyanobacteria detection method was verified by the traditional cell counting technique using a hemocytometer with an R² value of 0.993. It was shown that the limit of quantification of this microflow cytometry platform can be as low as 5 cells/mL for Microcystis aeruginosa, 400-fold lower than the Alert Level 1 (2000 cells/mL) set by the World Health Organization (WHO). Furthermore, the decreased detection limit may facilitate the future characterization of cyanobacterial bloom formation to better provide authorities with ample time to take the appropriate actions to mitigate human risk from these potentially hazardous blooms. Full article

Micro-devices that use electric fields to trap, analyze and inactivate micro-organisms vary in concept, design and application. The application of electric fields to manipulate and inactivate bacteria and single-celled organisms has been described extensively in the literature. By contrast, the effect of such fields on viruses is not well understood. This review explores the possibility of using existing methods for manipulating and inactivating larger viruses and bacteria, for smaller viruses, such as SARS-CoV-2. It also provides an overview of the theoretical background. The findings may be used to implement new ideas and frame experimental parameters that optimize the manipulation, sampling and inactivation of SARS-CoV-2 electrically. Full article

3D-printing is poised to enable remarkable advances in a variety of fields, such as artificial muscles, prosthetics, biomedical diagnostics, biofuel cells, flexible electronics, and military logistics. The advantages of automated monolithic fabrication are particularly attractive for complex embedded microfluidics in a wide range of applications. However, before this promise can be fulfilled, the basic problem of removal of sacrificial material from embedded microchannels must be solved. The presented work is an experimental proof of principle of a novel technique for clearance of sacrificial material from embedded microchannels in 3D-printed microfluidics. The technique demonstrates consistent performance (~40–75% clearance) in microchannels with printed width of ~200 µm and above. The presented technique is thus an important enabling tool in achieving the promise of 3D printing in microfluidics and its wide range of applications. Full article

Low-cost diagnostic tools for point-of-care immunoassays, such as the paper-based enzyme-linked immunoassay (ELISA), have become increasingly important, especially so in the recent COVID-19 pandemic. ELISA is the gold-standard antibody/antigen sensing method. This paper reports an easy-to-fabricate nitrocellulose (NC) paper plate, coupled with a desktop scanner for ELISA, which provides a higher protein immobilization efficiency than the conventional cellulose paper-based ELISA platforms. The experiments were performed using spiked samples for the direct ELISA of rabbit IgG with a limit of detection (LOD) of 1.016 μg/mL, in a measurement range of 10 ng/mL to 1 mg/mL, and for the sandwich ELISA of sperm protein (SP-10) with an LOD of 88.8 ng/mL, in a measurement range of 1 ng/mL to 100 μg/mL. The described fabrication method, based on laser-cutting, is a highly flexible one-step laser micromachining process, which enables the rapid production of low-cost NC paper-based multi-well plates with different sizes for the ELISA measurements. Full article

In this study, we propose a microfluidic organoid-trapping device used to immobilize human intestinal organoids and apply fluidic stimuli to them. The proposed device has a microchannel with a trapping region with wall gaps between the channel walls and the bottom surface, and a constriction to clog the organoids in the channel. Since the introduced culture medium escapes from the gap, organoids can be cultured without excessive deformation by hydrostatic pressure. Owing to the characteristics of the organoid-trapping device, we succeeded in trapping human intestinal organoids in the channel. Furthermore, to demonstrate the applicability of the device for culturing intestinal organoids, we induced organoid fusion to form large organoids by aligning the organoids in the channel and applying fluidic shear stress to the organoids to regulate their surface structures. Therefore, we believe that organoid-trapping devices will be useful for investigating organoids aligned or loaded with fluidic stimulation. Full article

Genetically-modified monoclonal cell lines are currently used for monoclonal antibody (mAbs) production and drug development. The isolation of single transformed cells is the main hindrance in the generation of monoclonal lines. Although the conventional limiting dilution method is time-consuming, laborious, and skill-intensive, high-end approaches such as fluorescence-activated cell sorting (FACS) are less accessible to general laboratories. Here, we report a bench-top approach for isolating single Chinese hamster ovary (CHO) cells using an adapted version of a simple microwell-based microfluidic (MBM) device previously reported by our group. After loading the cell suspension to the device, the electrostatically trapped cells can be viewed under a microscope and transferred using a micropipette for further clone establishment. Compared to the conventional method, the invented approach provided a 4.7-fold increase in the number of single cells isolated per round of cell loading and demonstrated a 1.9-fold decrease in total performing time. Additionally, the percentage of correct single-cell identifications was significantly improved, especially in novice testers, suggesting a reduced skill barrier in performing the task. This novel approach could serve as a simple, affordable, efficient, and less skill-intensive alternative to the conventional single-cell isolation for monoclonal cell line establishment. Full article

This paper reports a tunable optofluidic splitter in which the incident light is split via refraction and reflection at the interface between two laminar flows in a microchannel but with different refractive indices. A Y-junction microchannel is used to demonstrate the continuous tuning of the splitting ratio of optical power by smooth adjustment of the ratio of two flow rates. In addition, it has achieved the tuning of split angle from 5° to 19° by the control of the refractive index contrast. The dynamic response gives a fastest switching frequency of 1.67 Hz between the “wave-guiding” and “splitting” status. Full article

The microfluidic device (MFD) with a glass–PDMS–glass (G-P-G) structure is of interest for a wide range of applications. However, G-P-G MFD fabrication with an ultra-thin PDMS film (especially thickness less than 200 μm) is still a big challenge because the ultra-thin PDMS film is easily deformed, curled, and damaged during demolding and transferring. This study aimed to report a thickness-controllable and low-cost fabrication process of the G-P-G MFD with an ultra-thin PDMS film based on a flexible mold peel-off process. A patterned photoresist layer was deposited on a polyethylene terephthalate (PET) film to fabricate a flexible mold that could be demolded softly to achieve a rigid structure of the glass–PDMS film. The thickness of ultra-thin patterned PDMS could reach less than 50 μm without damage to the PDMS film. The MFD showcased the excellent property of water evaporation inhibition (water loss < 10%) during PCR thermal cycling because of the ultra-thin PDMS film. Its low-cost fabrication process and excellent water evaporation inhibition present extremely high prospects for digital PCR application. Full article

The virtual impactor, as an atmospheric particle classification chip, provides scientific guidance for identifying the characteristics of particle composition. Most of the studies related to virtual impactors focus on their size structure design, and the effect of temperature in relation to the dynamic viscosity on the cut−off diameter is rarely considered. In this paper, a new method that can reduce the cut−off particle size without increasing the pressure drop is proposed. Based on COMSOL numerical simulations, a new ultra−low temperature virtual impactor with a cut−off diameter of 2.5 μm was designed. A theoretical analysis and numerical simulation of the relationship between temperature and the performance of the virtual impactor were carried out based on the relationship between temperature and dynamic viscosity. The effects of inlet flow rate (Q), major flow channel width (S), minor flow channel width (L) and split ratio (r) on the performance of the virtual impactor were analyzed. The collection efficiency curves were plotted based on the separation effect of the new virtual impactor on different particle sizes. It was found that the new ultra−low temperature approach reduced the PM2.5 cut−off diameter by 19% compared to the conventional virtual impactor, slightly better than the effect of passing in sheath gas. Meanwhile, the low temperature weakens Brownian motion of the particles, thus reducing the wall loss. In the future, this approach can be applied to nanoparticle virtual impactors to solve the problem of their large pressure drop. Full article

Rapid prototyping methods enable the widespread adoption of microfluidic technologies by empowering end-users from non-engineering disciplines to make devices using processes that are rapid, simple and inexpensive. In this work, we developed a liquid molding technique to create silicone/PDMS microfluidic devices by replica molding. To construct a liquid mold, we use inexpensive adhesive-backed paper, an acetate backing sheet, and an off-the-shelf digital cutter to create paper molds, which we then wet with predetermined amounts of water. Due to the immiscibility of water and PDMS, mold patterns can be effectively transferred onto PDMS similarly to solid molds. We demonstrate the feasibility of these wet paper molds for the fabrication of PDMS microfluidic devices and assess the influence of various process parameters on device yield and quality. This method possesses some distinct benefits compared to conventional techniques such as photolithography and 3D printing. First, we demonstrate that the shape of a channel’s cross-section may be altered from rectangular to semicircular by merely modifying the wetting parameters. Second, we illustrate how electrical impedance can be utilized as a marker for inspecting mold quality and identifying defects in a non-invasive manner without using visual tools such as microscopes or cameras. As a proof-of-concept device, we created a microfluidic T-junction droplet generator to produce water droplets in mineral oil ranging in size from 1.2 µL to 75 µL. We feel that this technology is an excellent addition to the microfluidic rapid prototyping toolbox and will find several applications in biological research. Full article

Expansion of the microfluidics field dictates the necessity to constantly improve technologies used to produce such systems. One of the approaches which are used more and more is femtosecond (fs) direct laser writing (DLW). The subtractive model of DLW allows for directly producing microfluidic channels via ablation in an extremely simple and cost-effective manner. However, channel surface roughens are always a concern when direct fs ablation is used, as it normally yields an RMS value in the range of a few µm. One solution to improve it is the usage of fs bursts. Thus, in this work, we show how fs burst mode ablation can be optimized to achieve sub-µm surface roughness in glass channel fabrication. It is done without compromising on manufacturing throughput. Furthermore, we show that a simple and cost-effective channel sealing methodology of thermal bonding can be employed. Together, it allows for production functional Tesla valves, which are tested. Demonstrated capabilities are discussed. Full article

The catalysts employed in catalytic reactors greatly affect the reaction efficiency of the reaction system and the reactor’s performance. This work presents a rapid comparative study on three kinds of Fe-based materials integrated into an optofluidic Fenton reactor for water treatment. The Fe-based sheets (FeSiB, FeNbCuSiB, and FeNi) were respectively implanted into the reaction chamber to degrade the organic dyes with the assistance of H₂O₂. In the experiment, by adjusting the hydrogen peroxide concentration, flow rate, and light irradiation, the applicable conditions of the Fe-based materials for the dye degradation could be evaluated quickly to explore the optimal design of the Fenton reaction system. The results indicated that FeNi (1j85) exhibits excellent degradability in the microreactor, the reaction rate can reach 23.4%/s at the flow rate of 330 μL/min, but its weak corrosion resistance was definitely demonstrated. Although the initial degradability of the microreactor by using FeNbCuSiB (1k107) was not as good as that of 1j85, it increased after being reused several times instead, and the degradation efficiency reached >98% after being reused five times. However, the FeSiB (1k101) material shows the worst degradability and recycling. Therefore, in contrast, 1k107 has the greatest potential to be used in Fenton reactors for practical water treatment. Full article

A numerical simulation model of embedded liquid microchannels for cooling 3D multi-core chips is established. For the thermal management problem when the operating power of a chip changes dynamically, an intelligent method combining BP neural network and genetic algorithm is used for distribution optimization of coolant flow under the condition with a fixed total mass flow rate. Firstly, a sample point dataset containing temperature field information is obtained by numerical calculation of convective heat transfer, and the constructed BP neural network is trained using these data. The “working condition–flow distribution–temperature” mapping relationship is predicted by the BP neural network. The genetic algorithm is further used to optimize the optimal flow distribution strategy to adapt to the dynamic change of power. Compared with the commonly used uniform flow distribution method, the intelligently optimized nonuniform flow distribution method can further reduce the temperature of the chip and improve the temperature uniformity of the chip. Full article