Oocytes have long been used as a fundamental biological model in bioengineering research such as gene expression analysis, electrophysiological measurements, and drug screening. However, current methods mainly focus on single-microscope setups, which limits the efficient manipulation of multiple oocytes across spatially separated worksites. This study presents a dual vision-equipped microfluidic chip for the manipulation of multiple oocytes between different worksites. The microfluidic chip is equipped with two miniature cameras and then installed on a robotic manipulator. One miniature camera is used to track the position of oocytes in the microfluidic chip. The vision position of the multiple oocytes is utilized to control the flow. The results showed that multiple objects were successfully separated and released in sequence only using the hydrodynamic flow focusing effect. Moreover, a well port is designed to trap single oocytes to deal with the unseparated case of neighboring oocytes in the microchannel based on the vision information. Subsequently, the other camera is installed on the top of the tip part and utilized to detect the single object picking-placing position. Finally, we demonstrate that the dual vision-equipped microfluidic chip on-robot can pick, transport, and place multiple oocytes between different well chip areas. The proposed method has application potential in oocyte biomedical engineering.

The rheotactic behavior of spermatozoa in an outward radial flow was studied using a combination of theoretical and experimental approaches. Simulations suggested that at moderate flow rates, spermatozoa exhibited directed migration towards the flow origin, while at higher flow rates, the spermatozoa were predicted to display rotational rheotaxis, circling the origin while gradually moving inward, delaying their arrival at the center. Leveraging these findings, we developed a microfluidic device combining radial flow with strategically placed contracted pathways for efficient sperm selection, the sperm unidirectional navigation chip (SUN chip). This platform processed raw bovine semen to isolate highly motile (up to 98% motility) and vital spermatozoa suitable for assisted reproductive technologies. The yield of motile sperm retrieval for the SUN chip is ∼30%, and the cells in the processed population were 50% faster on average compared to raw semen. Our results demonstrate the potential of radial flow microfluidics to enhance sperm selection processes and can be used to investigate other microswimmer dynamics.

This study introduces a robust selective on-chip DNA synthesis platform utilizing an electric field-assisted polymerase chain reaction (E-PCR) system based on a microelectrode array. We demonstrate the use of electric field manipulation for the selective immobilization of DNA and enzyme-mediated single-stranded DNA synthesis, achieving precise spatial control at the microscale. By optimizing electric field patterns, we achieved high-efficiency synthesis and addressed critical challenges in electrochemical stability and DNA integrity. The E-PCR system enables selective DNA synthesis, length-controlled synthesis, and sensitive nucleic acid detection. Our findings provide a scalable foundation for high-throughput enzymatic synthesis, offering significant potential for DNA data storage, synthetic biology, and molecular diagnostics.

Human epidermal growth factor receptor 2 (HER2) is a key biomarker in breast cancer diagnosis, prognosis, and therapeutic decision-making. While its determination is traditionally based on tissue biopsy, serum-based testing is a simpler and less invasive alternative. However, its implementation remains challenging due to matrix effects and interference from high-abundance proteins, leading to insufficient sensitivity and specificity. To overcome these limitations, this work reports the development and optimization of a novel microfluidic immunoassay for HER2 detection in human serum, incorporating an innovative sample pretreatment step using a dual ionic liquid aqueous biphasic system (IL-IL-ABS). Under optimized conditions, the ABS composed of tri(isobutyl)methylphosphonium tosylate ([Pi(444)1][TsO]) and cholinium dihydrogenphosphate ([Ch][H2PO4]) effectively depleted high-abundance proteins (up to 100% removal) while selectively extracting 97% of HER2 into a separate aqueous phase. HER2 detection was performed in a protein G bead-based microfluidic device, demonstrating robust performance across diverse matrices. The limit of detection in ABS-pretreated serum (14.06 ng mL-1) was significantly improved compared to direct serum analysis (24.33 ng mL-1) and PBS (18.99 ng mL-1). By integrating advanced sample pretreatment with a sensitive microfluidic immunoassay, this work enhances the accuracy of HER2 quantification in complex biological samples, offering a promising approach for improved breast cancer detection and point-of-care applications.

The lymphatic system—integral to fluid balance, immune surveillance, and lipid absorption—is frequently overlooked despite its vital roles. Traditional research modalities, including static two-dimensional cultures and animal models, have illuminated key molecular and cellular features but fall short in recapitulating human lymphatic function, due to limited physiological relevance, throughput, and mechanobiological complexity. Recent advances in microfluidic organ-on-a-chip systems offer biomimetic platforms that integrate three-dimensional architecture, fluid flow, and biomechanical stimuli alongside human lymphatic endothelial and supporting cells. These lymphatics-on-a-chip constructs faithfully reproduce dynamic behaviors such as fluid drainage, junction remodeling, and cell trafficking under physiological and pathological responses. This review highlights the foundational lymphatic biology and engineering principles behind these devices, their capacity for disease modeling and drug testing, and their potential to drive future innovation through induced pluripotent stem cell integration, organ-specific customization, and computational modeling. Merging bioengineering, cell biology, and machine learning, lymphatic microphysiological systems stand poised to significantly expand our understanding and treatment of lymphatic-related disorders.

Magnetic fields can be used to control the process of magnetic fluid dispersion in microfluidics. Here we demonstrate a method for generating magnetic fluid droplets of a given size and shape in a microfluidic chip with a flow-focusing configuration under the influence of a transverse magnetic field. The results of the study show that a magnetic field can have a significant impact on ferrofluid dispersion. At low volumetric flow rates of the continuous phase, magnetic force plays a primary role in ferrofluid dispersion due to disturbances on the magnetic fluid surface. Furthermore, the droplet diameter decreases with increasing magnetic field Dd ∼ H-1/3. At high volumetric flow rates of the continuous phase, shear stress plays a primary role in ferrofluid dispersion. In this case, the droplet diameter depends on the continuous phase velocity according to the law Dd ∼ u-1. The magnetic field and the volumetric flow rate of the continuous phase affect not only the liquid dispersion process but also the coalescence and deformation of droplets. The pressure gradient in the channel and the magnetic field contribute to droplet deformation. The thickness of the deformed droplet decreases with increasing volumetric flow rate according to the law l ∼ Q-1. The non-uniformity of the velocity field distribution over the channel thickness is responsible for droplet coalescence. The magnetic field, on the contrary, prevents droplet coalescence. These findings open up new possibilities for generating soft magnetic robots of a given size and shape.

The paper demonstrates an adaptation of a Prusa Mini+ 3D printer through the integration of 3D printed modules, creating a system capable of producing varied droplets from multiple Eppendorf tubes. Building upon our previous model, this system enhances calibration methodology enabling any fused deposition modeling (FDM) printer to produce mono-disperse droplets (coefficient of variance (CV%) <2% for train of 100 droplets) with 6900 assays per hour rate. The cost of the developed system is 85% lower than that of existing droplet generation solutions on the market, and 30% more economical than the previous iteration of the system. Additionally, the system's utility in quantification of agglutination assays is highlighted using image analysis, capable of distinguishing between agglutinated and non-agglutinated samples. By offering significant savings and ease of use, this system aims to lower the barriers to entry for microfluidic research, potentially broadening the scope of scientific exploration and application in this field.

Electrical pulse stimulation (EPS) is used to replicate motor neuron activation in muscle tissues, enabling in vitro studies of muscle contraction. However, both custom-built and commercial existing EPS systems often suffer from significant limitations, including limited scalability, high cost, and lack of flexibility for experimental adaptation. This work presents the Myo-MOVES platform, a practical solution for stimulating 3D skeletal muscle tissues. The device has been designed as an intuitive EPS system consisting of two main components: a selector and a stimulator that adapts to commercial 24-well culture plates. The Myo-MOVES selector enables targeted stimulation of single or multiple wells, while the stimulator delivers electrical signals via graphite electrodes to the plate containing 3D skeletal muscle samples. The Myo-MOVES platform was technically validated and employed as a proof of concept to investigate sarcolemmal damage induced by muscle contraction in Duchenne muscular dystrophy (DMD) 3D skeletal muscle tissues. Taking advantage of the versatility of the device, we validated Myo-MOVES through the assessment of force generation in DMD engineered muscle tissues and the detection of contraction-induced sarcolemmal damage via Evans blue dye uptake and the release of creatine kinase (CK), the gold standard marker of muscle damage. These findings demonstrate the feasibility of using Myo-MOVES to induce and study functionally relevant disease phenotypes in DMD 3D skeletal muscle tissues. These results highlight the system's potential as a valuable tool for future applications in the field of 3D skeletal muscle tissue engineering, including drug screening and the study of DMD therapies and other muscular diseases.

Polydimethylsiloxane (PDMS) chips are still the workhorses of academic microfluidics. Their production requires the fabrication of moulds, commonly produced using clean-room technologies. Light-based 3D printing and in particular, vat photopolymerization, material jetting and two-photon polymerization are rising techniques for the fabrication of moulds for PDMS replication, thanks to their accessibility, fast prototyping time, and improving resolution. Here, we are first reviewing the possibility opened by 3D printing for soft lithography, with a focus on mould designs. Then, inhibition of PDMS curing by photosensitive resins will be discussed as the main technical hurdle of 3D printed moulds. Fortunately, mould post-treatments are efficient solutions to eliminate this curing inhibition, which we gathered in a large database of post-treatment protocols from the literature.

DNA methylation is a crucial epigenetic modification used as a biomarker for early cancer progression. However, existing methods for DNA methylation analysis are complex, time-consuming, and prone to DNA degradation. This work demonstrates selective capture of unmethylated DNAs using ZnO nanowires without chemical or biological modifications, thereby concentrating methylated DNA, particularly those with high methylation levels that can predict cancer risk. We observe varying affinities between methylated and unmethylated DNA on ZnO nanowires, which may be influenced by differences in hydrogen bonding strength, potentially related to the effects of methylation on DNA strand behavior, including self-aggregation and stretching inhibition. As a result, the nanowire-based microfluidic device effectively collects unmethylated DNA, leading to a significantly increased ratio of methylated to unmethylated DNA, particularly for collecting low-concentration methylated DNA. This simplified microfluidic device, composed of ZnO nanowires, enables direct separation of specific methylated DNA, offering a potential approach for DNA methylation mapping in clinical disease diagnostics.