Extracellular vesicles (EVs) are emerging as promising candidates in therapeutic applications due to their unique ability to mediate intercellular communication and deliver biological cargo. With increasing interest in EV‐based therapies, the development of scalable, cost‐effective and regulatory‐compliant production methods is critical. Microfluidic platforms offer transformative potential in EV manufacturing, providing precise control over production conditions, enhanced purity and seamless integration with quality control systems. This review highlights the advantages of microfluidic technologies in EV production, including fine‐tuning of shear stress to optimise yield, advanced purification strategies that achieve high recovery and purity, and on‐chip capabilities for EV loading and surface modification. Key challenges such as scaling up production while maintaining sterility, controlling EV release after immunoaffinity capture, and addressing clogging and fouling in microfluidic devices are discussed alongside emerging solutions. Additionally, the integration of AI‐driven automation and real‐time monitoring, as well as personalised EV manufacturing, is explored as pivotal innovations. Future directions emphasise the potential of combining size‐ and affinity‐based methods for EV isolation and aligning microfluidic technologies with regulatory requirements to accelerate clinical translation. Therefore, we believe microfluidics platforms for EV isolation hold immense potential to redefine EV manufacturing by enabling scalable, reproducible and high‐quality production systems essential for therapeutic applications.

Mixing fluid samples or reactants is a paramount function in the fields of micro total analysis system (μTAS) and microchemical processing. However, rapid and efficient fluid mixing is difficult to achieve inside microchannels because of the difficulty of diffusive mass transfer in the laminar regime of the typical microfluidic flows. It has been well recorded that the mixing efficiency can be boosted by migrating from two-dimensional (2D) to three-dimensional (3D) geometries. Although several 3D chaotic mixers have been designed, most of them offer a high mixing efficiency only in a very limited range of Reynolds numbers (Re). In this work, we developed a 3D fine-threaded lemniscate-shaped micromixer whose maximum numerical and empirical efficiency is around 97% and 93%, respectively, and maintains its high performance (i.e., >90%) over a wide range of 1 < Re < 1000 which meets the requirements of both the μTAS and microchemical process applications. The 3D micromixer was designed based on two distinct mixing strategies, namely, the inducing of chaotic advection by the presence of Dean flow and diffusive mixing through thread-like grooves around the curved body of the mixers. First, a set of numerical simulations was performed to study the physics of the flow and to determine the essential geometrical parameters of the mixers. Second, a simple and cost-effective method was exploited to fabricate the convoluted structure of the micromixers through the removal of a 3D-printed wax structure from a block of cured polydimethylsiloxane. Finally, the fabricated mixers with different threads were tested using a fluorescent microscope demonstrating a good agreement with the results of the numerical simulation. We envisage that the strategy used in this work would expand the scope of the micromixer technology by broadening the range of efficient working flow rate and providing an easy way to the fabrication of 3D convoluted microstructures.

Cancer-derived small extracellular vesicles (sEVs) are specific subgroups of lipid bilayer vesicles secreted from cancer cells to the extracellular environment. They carry distinct biomolecules (e.g., proteins, lipids and nucleic acids) from their parent cancer cells. Therefore, the analysis of cancer-derived sEVs can provide valuable information for cancer diagnosis. However, the use of cancer-derived sEVs in clinics is still limited due to their small size, low amounts in circulating fluids, and heterogeneous molecular features, making their isolation and analysis challenging. Recently, microfluidic technology has gained great attention for its ability to isolate sEVs in minimal volume. In addition, microfluidics allows the isolation and detection of sEVs to be integrated into a single device, offering new opportunities for clinical application. Among various detection techniques, surface-enhanced Raman scattering (SERS) has emerged as a promising candidate for integrating with microfluidic devices due to its ultra-sensitivity, stability, rapid readout, and multiplexing capability. In this tutorial review, we start with the design of microfluidics devices for isolation of sEVs and introduce the key factors to be considered for the design, and then discuss the integration of SERS and microfluidic devices by providing descriptive examples of the currently developed platforms. Lastly, we discuss the current limitations and provide our insights for utilising integrated SERS-microfluidics to isolate and analyse cancer-derived sEVs in clinical settings.