A microfluidic method for passive trapping of sperms in microstructures.

Sperm motility is a prerequisite for male fertility. Enhancing the concentration of motile sperms in assisted reproductive technologies - for human and animal reproduction - is typically achieved through aggressive methods such as centrifugation. Here, we propose a passive technique for the amplification of motile sperm concentration, with no externally imposed forces or flows. The technique is based on the disparity between probability rates, for motile cells, of entering and escaping from complex structures. The effectiveness of the technique is demonstrated in microfluidic experiments with microstructured devices, comparing the trapping power in different geometries. In these micro-traps, we observe an enhancement of cells' concentration close to 10, with a contrast between motile and non-motile cells increased by a similar factor. Simulations of suitable interacting model sperms in realistic geometries reproduce quantitatively the experimental results, extend the range of observations and highlight the components that are key to the optimal trap design.

Deciphering Hydrodynamic and Drug-Resistant Behaviors of Metastatic EMT Breast Cancer Cells Moving in a Constricted Microcapillary.

Epithelial to mesenchymal transition (EMT) induces cell migration, invasion, and drug resistance, and consequently, contributes to cancer metastasis and disease aggressiveness. This study attempted to address crucial biological parameters to correlate EMT and drug-treated cancer cells traversing through microcapillaries, reminiscent of metastatic conditions. MDA-MB-468 breast cancer cells induced to undergo EMT by treatment with 20 ng/mL of epidermal growth factor (EGF) were initially passed through several blockages and then through a constricted microchannel, mimicking the flow of invasive metastatic cells through constricted blood microcapillaries. EMT cells acquired enhanced migratory properties and retained 50% viability, even after migration through wells 10-15 µm in size and a constricted passage of 7 µm and 150 µm in length at a constant flow rate of 50 µL/h. The hydrodynamic properties revealed cellular deformation with a deformation index, average transit velocity, and entry time of 2.45, 12.3 mm/s, and 31,000 µs, respectively for a cell of average diameter 19 µm passing through one of the 7 µm constricted sections. Interestingly, cells collected at the channel outlet regained epithelial character, undergoing reverse transition (mesenchymal to epithelial transition, MET) in the absence of EGF. Remarkably, real-time polymerase chain reaction (PCR) analysis confirmed increases of 2- and 2.7-fold in the vimentin and fibronectin expression in EMT cells, respectively; however, their expression reduced to basal level in the MET cells. A scratch assay revealed the pronounced migratory nature of EMT cells compared with MET cells. Furthermore, the number of colonies formed from EMT cells and paclitaxel-treated EMT cells after passing through a constriction were found to be 95 ± 10 and 79 ± 4, respectively, confirming that the EMT cells were more drug resistant with a concomitant two-fold higher expression of the multi-drug resistance (MDR1) gene. Our results highlight the hydrodynamic and drug-evading properties of cells that have undergone an EMT, when passed through a constricted microcapillary that mimics their journey in blood circulation.

Understanding flow dynamics, viability and metastatic potency of cervical cancer (HeLa) cells through constricted microchannel.

To understand the burgeoning challenges of metastasis, a microchannel of 35 µm diameter, constricted to 7 µm for a distance of 200 µm in a total length of 3 mm, was designed and fabricated using a mask aligner made of polydimethylsiloxane (PDMS) to mimic in vivo capillaries. A thin glass cover-slide was mounted on top to monitor the motion of single or aggregated malignant HeLa cells (size 17-30 µm) microscopically through the constricted microchannel at a constant flow rate of 30 µl/h. Quantitative deconvolution of high-speed videographs of a single cell of 30 µm revealed cellular deformation while passing through constriction, having elongation index, average transit velocity and entry time of 2.67, 18 mm/s and 5.1 ms, respectively. Morphological analysis of live and apoptotic cells by dual staining with Acridine Orange/Ethidium Bromide demonstrated retention of a significant viable cell population after exit through the constriction and a viability index of 50% was quantified by dye exclusion assay. The cumulative data for microfluidic parameters, morphology and relevant metastatic MMP2 gene expression efficiency measured by real-time polymerase chain reaction revealed retention of virulence potency that could possibly cause metastasis, would be beneficial in developing futuristic MEMS device for cancer theranostics.

Cross-stream migration of drops suspended in Poiseuille flow in the presence of an electric field.

The present study focuses on the cross-stream migration of a neutrally buoyant two-dimensional drop in a Poiseuille flow in a channel under the influence of an electric field. In the absence of an electric field, the important nondimensional parameters describing this problem are the viscosity ratio (λ) between the drop fluid and the surrounding medium, the ratio of drop diameter to channel height (a^{*}), and the capillary number (Ca). The influence of all these parameters on drop migration is investigated. It is observed that a large drop moves slowly as compared to a smaller drop, but attains a steady shape at the center line of the channel. The increase in value of the capillary number enhances the cross-stream migration rate, while the increase in viscosity ratio reduces the tendency of the drops to move towards the channel center line. The presence of an electric field introduces additional interfacial stresses at the drop interface, which in turn alters the dynamics observed in the absence of an electric field. Extensive computations are carried out to analyze the combined effect of the electric field and the shear flow on the cross-stream migration of the drop. The computational results for a perfect dielectric indicate that the droplet migration enhances in the presence of an electric field. The permittivity ratio (S) and the electric field strength (E) play major roles in drop migration and deformation. Computations using the leaky dielectric model also show that for certain combinations of electrical properties the drop undergoes immense elongation along the direction of the electric field. The conductivity ratio (R) is again a vital parameter in such a system of fluids. It is further observed that for certain conditions the leaky dielectric drops exhibit rotation together with translation.

Migration of a droplet in a cylindrical tube in the creeping flow regime.

The migration of a neutrally buoyant droplet in a tube containing another immiscible liquid is investigated numerically in the creeping flow regime. A fully developed velocity profile is imposed at the inlet of the tube. The interface between the two immiscible fluids is captured using a coupled level-set and volume-of-fluid approach. The deformation and breakup dynamics of the droplet are investigated in terms of three dimensionless parameters, namely, the ratio between the radius of the undeformed droplet and the radius of the capillary tube, the viscosity ratio between the dispersed and the continuous phases, and the capillary number that measures the relative importance of the viscous force over the surface tension force. It has been observed that the droplet, while traversing through the tube, either approaches a steady bulletlike shape or develops a prominent reentrant cavity at its rear. Depending on the initial droplet size, there exists a critical capillary number for every flow configuration beyond which the drop fails to maintain a steady shape and breaks into fragments. The deformation and breakup phenomena depend primarily on the droplet size, the viscosity ratio, and the capillary number. Special attention has been given to the case where the drop diameter is comparable with the tube diameter. A thorough computational study has been conducted to find the critical capillary number for a range of droplets of varied sizes suspended in systems having different viscosity ratios.