Neutrophil migration in tissues critically regulates the human immune response and can either play a protective role in host defense or cause health problems. Microfluidic chips are increasingly applied to study neutrophil migration, attributing to their advantages of low reagent consumption, stable chemical gradients, visualized cell chemotaxis monitoring, and quantification. Most chemotaxis chips suffered from low throughput and fussy cell separation operations. We here reported a novel and simple “sample in and chemotaxis out” method for rapid neutrophils isolation from a small amount of whole blood based on a simplified magnetic method, followed by a chemotaxis assay on a microfluidic chip (SC² chip) consisting of six cell migration units and six-cell arrangement areas. The advantages of the “sample in and chemotaxis out” method included: less reagent consumption (10 μL of blood + 1 μL of magnetic beads + 1 μL of lysis buffer); less time (5 min of cell isolation + 15 min of chemotaxis testing); no ultracentrifugation; more convenient; higher efficiency; high throughput. We have successfully validated the approach by measuring neutrophil chemotaxis to frequently-used chemoattractant (i.e., fMLP). The effects of D-glucose and mannitol on neutrophil chemotaxis were also analyzed. In addition, we demonstrated the effectiveness of this approach for testing clinical samples from diabetes mellitus type 2 (T2DM) patients. We found neutrophils’ migration speed was higher in the “well-control” T2DM than in the “poor-control” group. Pearson coefficient analysis further showed that the migration speed of T2DM was negatively correlated with physiological indicators, such as HbA1c (−0.44), triglyceride (−0.36), C-reactive protein (−0.28), and total cholesterol (−0.28). We are very confident that the developed “sample in and chemotaxis out” method was hoped to be an attractive model for analyzing the chemotaxis of healthy and disease-associated neutrophils. Full article

Owing to the complex and long-term treatment of foot wounds due to diabetes and the limited mobility of patients, advanced clinical surgery often uses wearable flexible devices for auxiliary treatment. Therefore, there is an urgent need for self-powered biomedical devices to reduce the extra weight. We have prepared an electrically stimulated MEMS (Micro Electromechanical System) electrode integrated with wearable OPV (Organic photovoltaic). The wearable OPV is constructed of a bio-affinity PET-ITO substrate and a hundred-nanometer organic layer. Under sunlight and near-infrared light irradiation, a voltage and current are supplied to the MEMS electrode to generate an exogenous lateral electric field directed to the center of the wound. The results of in vitro cell experiments and diabetic skin-relieving biological experiments showed the proliferation of skin fibroblasts and the expression of transforming growth factors increased, and the skin wounds of diabetic mouse healed faster. Our research provides new insights for the clinical treatment of diabetes. Full article

This study proposes a rapid and inexpensive thermocycler that enables rapid heating of samples using a thin glass chip and a cheap chip resistor to overcome the on-site diagnostic limitations of polymerase chain reaction (PCR). Microchip PCR devices have emerged to miniaturize conventional PCR systems and reduce operation time and cost. In general, PCR microchips require a thin-film heater fabricated through a semiconductor process, which is a complicated process, resulting in high costs. Therefore, this investigation substituted a general chip resistor for a thin-film heater. The proposed thermocycler consists of a compact glass microchip of 12.5 mm × 12.5 mm × 2 mm that could hold a 2 μL PCR sample and a surface-mounted chip resistor of 6432 size (6.4 mm × 3.2 mm). Improving heat transfer from the chip resistor heater to the PCR reaction chamber in the microchip was accomplished via the design and fabrication of a three-dimensional chip structure using selective laser-induced etching, a rapid prototyping technique that allowed to be embedded. The fabricated PCR microchip was combined with a thermistor temperature sensor, a blower fan, and a microcontroller. The assembled thermocycler could heat the sample at a maximum rate of 28.8 °C/s per second. When compared with a commercially available PCR apparatus running the same PCR protocol, the total PCR operating time with a DNA sample was reduced by about 20%. Full article

The ability to accurately quantify dielectrophoretic (DEP) force is critical in the development of high-efficiency microfluidic systems. This is the first reported work that combines a textile electrode-based DEP sensing system with deep learning in order to estimate the DEP forces invoked on microparticles. We demonstrate how our deep learning model can process micrographs of pearl chains of polystyrene (PS) microbeads to estimate the DEP forces experienced. Numerous images obtained from our experiments at varying input voltages were preprocessed and used to train three deep convolutional neural networks, namely AlexNet, MobileNetV2, and VGG19. The performances of all the models was tested for their validation accuracies. Models were also tested with adversarial images to evaluate performance in terms of classification accuracy and resilience as a result of noise, image blur, and contrast changes. The results indicated that our method is robust under unfavorable real-world settings, demonstrating that it can be used for the direct estimation of dielectrophoretic force in point-of-care settings. Full article

Neutrophil dysfunction is closely related to the pathophysiology of patients with diabetes mellitus, but existing immunoassays are difficult to implement in clinical applications, and neutrophil’s chemotaxis as a functional biomarker for diabetes mellitus prognostic remains largely unexplored. Herein, a novel microfluidic device consisted of four independent test units with four cell docking structures was developed to study the neutrophil chemotaxis, which allowed multiple cell migration observations under a single field of view (FOV) and guaranteed more reliable results. In vitro studies, the chemotaxis of healthy neutrophils to N-Formyl-Met-Leu-Phe (fMLP) gradient (0, 10, 100, and 1000 nM) was concentration-dependent. The distinct promotion or suppression in the chemotaxis of metformin or pravastatin pretreated cells were observed after exposure to 100 nM fMLP gradient, indicating the feasibility and efficiency of this novel microfluidic device for clinically relevant evaluation of neutrophil functional phenotype. Further, the chemotaxis of neutrophils pretreated with 25, 50, or 70 mM of glucose was quantitatively lower than that of the control groups (i.e., 5 mM normal serum level). Neutrophils exposed to highly concentrated advanced glycation end products (AGEs) (0.2, 0.5, or 1.0 μM; 0.13 μM normal serum AGEs level), a product of prolonged hyperglycemia, showed that the higher the AGEs concentration was, the weaker the migration speed became. Specifically, neutrophils exposed to high concentrations of glucose or AGEs also showed a stronger drifting along with the flow, further demonstrating the change of neutrophil chemotaxis. Interestingly, adding the N-benzyl-4-chloro-N-cyclohexylbenzamide (FPS-ZM1) (i.e., high-affinity RAGE inhibitor) into the migration medium with AGEs could hinder the binding between AGEs and AGE receptor (RAGE) located on the neutrophil, thereby keeping the normal chemotaxis of neutrophils than the ones incubated with AGEs alone. These results revealed the negative effects of high concentrations of glucose and AGEs on the neutrophil chemotaxis, suggesting that patients with diabetes should manage serum AGEs and also pay attention to blood glucose indexes. Overall, this novel microfluidic device could significantly characterize the chemotaxis of neutrophils and have the potential to be further improved into a tool for risk stratification of diabetes mellitus. Full article

The generation of droplets is one of the most critical steps in the droplet digital polymerase chain reaction (ddPCR) procedure. In this study, the mechanism of droplet formation in microchannel structure and factors affecting droplet formation were studied. The physical field of laminar two-phase flow level was used to simulate the process of droplet generation through microfluidic technology. The effect of the parameters including flow rate, surface tension, and viscosity on the generated droplet size were evaluated by the simulation. After that, the microfluidic chip that has the same dimension as the simulation was then, fabricated and evaluated. The chip was made by conventional SU-8 photolithography and injection molding. The accuracy of the simulation was validated by comparing the generated droplets in the real scenario with the simulation result. The relative error (RE) between experimentally measured droplet diameter and simulation results under different flow rate, viscosity, surface tension and contact angle was found less than 3.5%, 1.8%, 1.4%, and 1.2%, respectively. Besides, the coefficient of variation (CV) of the droplet diameter was less than 1%, which indicates the experimental droplet generation was of high stability and reliability. This study provides not only fundamental information for the design and experiment of droplet generation by microfluidic technology but also a reliable and efficient investigation method in the ddPCR field. Full article

Since being invented, droplet microfluidic technologies have been proven to be perfect tools for high-throughput chemical and biological functional screening applications, and they have been heavily studied and improved through the past two decades. Each droplet can be used as one single bioreactor to compartmentalize a big material or biological population, so millions of droplets can be individually screened based on demand, while the sorting function could extract the droplets of interest to a separate pool from the main droplet library. In this paper, we reviewed droplet detection and active sorting methods that are currently still being widely used for high-through screening applications in microfluidic systems, including the latest updates regarding each technology. We analyze and summarize the merits and drawbacks of each presented technology and conclude, with our perspectives, on future direction of development. Full article

The significant advancements within the electronics miniaturization field have shifted the scientific interest towards a new class of precision devices, namely microelectromechanical systems (MEMS). Specifically, MEMS refers to microscaled precision devices generally produced through micromachining techniques that combine mechanical and electrical components for fulfilling tasks normally carried out by macroscopic systems. Although their presence is found throughout all the aspects of daily life, recent years have witnessed countless research works involving the application of MEMS within the biomedical field, especially in drug synthesis and delivery, microsurgery, microtherapy, diagnostics and prevention, artificial organs, genome synthesis and sequencing, and cell manipulation and characterization. Their tremendous potential resides in the advantages offered by their reduced size, including ease of integration, lightweight, low power consumption, high resonance frequency, the possibility of integration with electrical or electronic circuits, reduced fabrication costs due to high mass production, and high accuracy, sensitivity, and throughput. In this context, this paper aims to provide an overview of MEMS technology by describing the main materials and fabrication techniques for manufacturing purposes and their most common biomedical applications, which have evolved in the past years. Full article