Viscoelastic microfluidics for enhanced separation resolution of submicron particles and extracellular vesicles.

Manipulation, focusing, and separation of submicron- and nanoparticles such as extracellular vesicles (EVs), viruses and bacteria have broad applications in disease diagnostics and therapeutics. Viscoelastic microfluidic technology emerges as a promising technique, and it shows an unparalleled capacity to manipulate and separate submicron particles in a high resolution based on the elastic effects of non-Newtonian mediums. The maximum particle separation resolution for the reported state-of-the-art viscoelastic microfluidics is around 200 nm. To further enhance the reseparation resolution, this work develops a viscoelastic microfluidic device that can achieve a finer separation resolution up to 100 nm, by optimising the operating conditions such as flow rate, flow rate ratio and polyethylene oxide (PEO) concentration. With these optimised conditions, we separated a ternary mixture of 100 nm, 200 nm and 500 nm polystyrene particles, with purities above 90%, 70% and 82%, respectively. Furthermore, we also applied the developed viscoelastic microfluidic device for the separation of cancer cell-secreted extracellular vesicles (EVs) into three different size groups. After single processing, the separation efficiencies for small EVs (sEVs, <150 nm), medium EVs (mEVs, 150-300 nm), and large EVs (>300 nm) were 86%, 80% and 50%, respectively. The enrichment factors for the three EV groups were 2.4, 1.1 and 1.3, respectively. Moreover, we observed an unexpected effect of high PEO concentrations (2000-5000 ppm) on the lateral migration of nanoparticles where nanoparticles of up to 50 nm surprisingly can migrate and concentrate at the middle of the microchannel. This simple and label-free viscoelastic microfluidic device possesses excellent potential for sorting submicron particles for various chemical, biological, medical and environmental applications.