In-flow measurement of cell-cell adhesion using oscillatory inertial microfluidics.

Multicellular clusters in circulation can exhibit a substantially different function and biomarker significance compared to individual cells. Notably, clusters of circulating tumor cells (CTCs) are much more effective initiators of metastasis than single CTCs, and correlate with worse patient prognoses. Measuring the cell-cell adhesion strength of CTC clusters is a critical step towards understanding their subsistence in the circulation and mechanism of elevated tumorigenicity. However, measuring cell-cell adhesion forces in flow is elusive using existing methods. Here, we report an oscillatory inertial microfluidics system which exerts a repeating fluidic force profile on suspended cell doublets to determine their cell-cell adhesion strength (Fs), without any biophysical modifications to the cell surface and physiological morphology. Using our system, we analyzed a large number (N > 500) of doublets from a patient-derived breast cancer CTC line. We discovered that the cell-cell adhesion strength of CTC doublets varied almost 20-fold between the weakly adhered (Fs < 28 nN) and strongly bound subpopulations (Fs > 542 nN). Our system can be used with other cancer or noncancer cells without restrictions, and may be used for rapid screening of drugs aiming to disrupt the highly-metastatic CTC clusters in circulation.

Inertial Focusing of Microparticles in Curvilinear Microchannels.

A passive, continuous and size-dependent focusing technique enabled by "inertial microfluidics", which takes advantage of hydrodynamic forces, is implemented in this study to focus microparticles. The objective is to analyse the decoupling effects of inertial forces and Dean drag forces on microparticles of different sizes in curvilinear microchannels with inner radius of 800 µm and curvature angle of 280°, which have not been considered in the literature related to inertial microfluidics. This fundamental approach gives insight into the underlying physics of particle dynamics and offers continuous, high-throughput, label-free and parallelizable size-based particle separation. Our design allows the same footprint to be occupied as straight channels, which makes parallelization possible with optical detection integration. This feature is also useful for ultrahigh-throughput applications such as flow cytometers with the advantages of reduced cost and size. The focusing behaviour of 20, 15 and 10 µm fluorescent polystyrene microparticles was examined for different channel Reynolds numbers. Lateral and vertical particle migrations and the equilibrium positions of these particles were investigated in detail, which may lead to the design of novel microfluidic devices with high efficiency and high throughput for particle separation, rapid detection and diagnosis of circulating tumour cells with reduced cost.