Exosomes, molecular cargos secreted by almost all mammalian cells, are considered as promising biomarkers to identify many diseases including cancers. However, the small size of exosomes (30−200 nm) poses serious challenges in their isolation from complex media containing a variety of extracellular vesicles (EVs) of different sizes, especially in small sample volumes. Here we present a viscoelasticitybased microfluidic system to directly separate exosomes from cell culture media or serum in a continuous, size-dependent, and label-free manner. Using a small amount of biocompatible polymer as the additive in the media to control the viscoelastic forces exerted on EVs, we are able to achieve a high separation purity (>90%) and recovery (>80%) of exosomes. The proposed technique may serve as a versatile platform to facilitate exosome analyses in diverse biochemical applications.

Interaction with the pulmonary surfactant film, being the first line of host defense, represents the initial bio-nano interaction in the lungs. Such interaction determines the fate of the inhaled nanoparticles and their potential therapeutic or toxicological effect. Despite considerable progress in optimizing physicochemical properties of nanoparticles for improved delivery and targeting, the mechanisms by which inhaled nanoparticles interact with the pulmonary surfactant film are still largely unknown. Here, using combined in vitro and in silico methods, we show how hydrophobicity and surface charge of nanoparticles differentially regulate the translocation and interaction with the pulmonary surfactant film. While hydrophilic nanoparticles generally translocate quickly across the pulmonary surfactant film, a significant portion of hydrophobic nanoparticles are trapped by the surfactant film and encapsulated in lipid protrusions upon film compression. Our results support a novel model of pulmonary surfactant lipoprotein corona associated with inhaled nanoparticles of different physicochemical properties. Our data suggest that the study of pulmonary nanotoxicology and nanoparticle-based pulmonary drug delivery should consider this lipoprotein corona.

This work reports on a passive double spiral microfluidic device allowing rapid and label-free tumor cell separation and enrichment from diluted peripheral whole blood, by exploiting the size-dependent hydrodynamic forces. A numerical model is developed to simulate the Dean flow inside the curved geometry and to track the particle/cell trajectories, which is validated against the experimental observations and serves as a theoretical foundation for optimizing the operating conditions. Results from separating tumor cells (MCF-7 and Hela) spiked into whole blood indicate that 92.28% of blood cells and 96.77% of tumor cells are collected at the inner and the middle outlet, respectively, with 88.5% tumor recovery rate at a throughput of 3.33 6 10 7 cells min 21. We expect that this label-free microfluidic platform, driven by purely hydrodynamic forces, would have an impact on fundamental and clinical studies of circulating tumor cells.

a b s t r a c tMicrofluidic devices have been widely used since 1990s for diverse manipulations of particles (a general term of beads, cells, vesicles, drops, etc.) in a variety of applications. Compared to the active manipulation via an externally imposed force field, the passive manipulation of particles exploits the flow-induced intrinsic lift and/or drag to control particle motion with several advantages. Along this direction, inertial microfluidics has received tremendous interest in the past decade due to its capability to handle a large volume of samples at a high throughput. This inertial lift-based approach in Newtonian fluids, however, becomes ineffective and even fails for small particles and/or at low flow rates. Recent studies have demonstrated the potential of elastic lift in non-Newtonian fluids for manipulating particles with a much smaller size and over a much wider range of flow rates. The aim of this article is to provide an overview of the various passive manipulations, including focusing, separation, washing and stretching, of particles that have thus far been demonstrated in non-Newtonian microfluidics.

Viscoelasticity-induced particle migration has recently received increasing attention due to its ability to obtain high-quality focusing over a wide range of flow rates. However, its application is limited to low throughput regime since the particles can defocus as flow rate increases. Using an engineered carrier medium with constant and low viscosity and strong elasticity, the sample flow rates are improved to be one order of magnitude higher than those in existing studies. Utilizing differential focusing of particles of different sizes, here we present sheathless particle/cell separation in simple straight microchannels that possess excellent parallelizability for further throughput enhancement. The present method can be implemented over a wide range of particle/cell sizes and flow rates. We successfully separate small particles from larger particles, MCF-7 cells from red blood cells (RBCs), and Escherichia coli (E. coli) bacteria from RBCs in different straight microchannels. The proposed method could broaden the applications of viscoelastic microfluidic devices to particle/cell separation due to the enhanced sample throughput and simple channel design.Continuous manipulation and separation of particles and cells is important for a wide range of applications in biology, 1,2 medicine, 2,3 and industry. [4][5][6] Microfluidic systems have been proven to be promising tools for particle/cell manipulation with higher sensitivity and accuracy than their macroscale counterparts. The last decade has seen extensive development of microfluidic approaches for particle/cell manipulation that resort to immunocapture, 7 externally applied physical fields, [8][9][10][11][12][13][14][15][16][17][18] microfiltration, 19,20 gravitational sedimentation, 21 or deterministic lateral migration. 22,23 More recently, cross-streamline migration induced by the hydrodynamic effects of carrier media, such as inertia 24,25 and viscoelasticity, 26,27 has shown its promise for effective particle/cell manipulation without need of labeling and external force fields. Particles and cells can be separated based on the size-dependent nature of hydrodynamic forces. Briefly, the inertial lift scales as F a ∝ , where a is the particle diameter. There are several cell types of biological and clinical interest with separable size ranges: epithelial tumor cells (15-25 µm in diameter), blood cells (erythrocytes are 6-8 µm biconcave disks and peripheral blood lymphocytes are 7-10 µm in diameter), and bacteria (1-3 µm). Inertial migration in Newtonian fluids has been intensively studied and implemented in high-throughput label-free separation devices for cell separation. [28][29][30][31][32][33][34] Recently, particle migration induced by viscoelasticity has begun to attract increasing attention due to its simple focusing pattern and potential for achieving efficient focusing over a wide range of flow rates. 35,36 In a viscoelastic medium, elasticity coupled with non-negligible inertia will drive particles towards the channel centerline, whic...

Inertial microfluidics has emerged as an important tool for manipulating particles and cells. For a better design of inertial microfluidic devices, we conduct 3D direct numerical simulations (DNS) and experiments to determine the complicated dependence of focusing behaviour on the particle size, channel aspect ratio, and channel Reynolds number. We find that the well-known focusing of the particles at the two centers of the long channel walls occurs at a relatively low Reynolds number, whereas additional stable equilibrium positions emerge close to the short walls with increasing Reynolds number. Based on the numerically calculated trajectories of particles, we propose a two-stage particle migration which is consistent with experimental observations. We further present a general criterion to secure good focusing of particles for high flow rates. This work thus provides physical insight into the multiplex focusing of particles in rectangular microchannels with different geometries and Reynolds numbers, and paves the way for efficiently designing inertial microfluidic devices.

Joule heating effects on electroosmotic flow in insulator-based dielectrophoresisInsulator-based dielectrophoresis (iDEP) is an emerging technology that has been successfully used to manipulate a variety of particles in microfluidic devices. However, due to the locally amplified electric field around the in-channel insulator, Joule heating often becomes an unavoidable issue that may disturb the electroosmotic flow and affect the particle motion. This work presents the first experimental study of Joule heating effects on electroosmotic flow in a typical iDEP device, e.g. a constriction microchannel, under DC-biased AC voltages. A numerical model is also developed to simulate the observed flow pattern by solving the coupled electric, energy, and fluid equations in a simplified two-dimensional geometry. It is observed that depending on the magnitude of the DC voltage, a pair of counter-rotating fluid circulations can occur at either the downstream end alone or each end of the channel constriction. Moreover, the pair at the downstream end appears larger in size than that at the upstream end due to DC electroosmotic flow. These fluid circulations, which are reasonably simulated by the numerical model, form as a result of the action of the electric field on Joule heatinginduced fluid inhomogeneities in the constriction region.

Focusing particles into a tight stream is usually a necessary step prior to separating and sorting them. We present herein a proof-of-concept experiment of a novel particle focusing technique in DC electrokinetic flow through a planar serpentine microchannel. This focusing stems from the cross-stream dielectrophoretic motion induced within the channel turns. The observed particle focusing behavior is consistent with the predicted particle trajectories from a numerical modeling.