Microfluidic droplet arrays potentially allow to carry out assays involving complex workflows at the single-cell level. In recent developments [e.g. (Jeong et al. in Lab Chip 16:1698, 2016)], each droplet of the array can be addressed individually thanks to on-chip pneumatic valves. In this experimental and theoretical work, we investigate the multiple interaction regimes between one or several droplets and a pneumatic valve. In particular, two main trapping modes are characterized, with very distinct flow features. We quantify the potentially huge increase of hydrodynamic resistance induced by the droplet/valve interaction. Finally, we rationalize the main transitions between trapped and non-trapped regimes through a generic theoretical model of the balance between capillary, hydrodynamic and elastic forces.

Background:In vitro maturation of immature testicular tissue (ITT) cryopreserved for fertility preservation is a promising fertility restoration strategy. Organotypic tissue culture proved successful in mice, leading to live births. In larger mammals, including humans, efficiently reproducing spermatogenesis ex vivo remains challenging. With advances in biomaterials technology, culture systems are becoming more complex to better mimic in vivo conditions. Along with improving culture media components, optimizing physical culture conditions (e.g., tissue perfusion, oxygen diffusion) also needs to be considered. Recent studies in mice showed that by using silicone-based hybrid culture systems, the efficiency of spermatogenesis can be improved. Such systems have not been reported for ITT of large mammals. Methods: Four different organotypic tissue culture systems were compared: static i.e., polytetrafluoroethylene membrane inserts (OT), agarose gel (AG) and agarose gel with polydimethylsiloxane chamber (AGPC), and dynamic i.e., microfluidic (MF). OT served as control. Porcine ITT fragments were cultured over a 30-day period using a single culture medium. Analyses were performed at days (d) 0, 5, 10, 20 and 30. Seminiferous tubule (ST) integrity, diameters, and tissue core integrity were evaluated on histology. Immunohistochemistry was used to identify germ cells (PGP9.5, VASA, SYCP3, CREM), somatic cells (SOX9, INSL3) and proliferating cells (Ki67), and to assess oxidative stress (MDA) and apoptosis (C-Caspase3). Testosterone was measured in supernatants using ELISA. Results: ITT fragments survived and grew in all systems. ST diameters, and Sertoli cell (SOX9) numbers increased, meiotic (SYCP3) and post-meiotic (CREM) germ cells were generated, and testosterone was secreted. When compared to control (OT), significantly larger STs (d10 through d30), better tissue core integrity (d5 through d20), higher numbers of undifferentiated spermatogonia (d30), meiotic and post-meiotic germ cells (SYCP3: d20 and 30, CREM: d20) were observed in the AGPC system. Apoptosis, lipid peroxidation (MDA), ST integrity, proliferating germ cell (Ki67/VASA) numbers, Leydig cell (INSL3) numbers and testosterone levels were not significantly different between systems. Conclusions: Using a modified culture system (AGPC), germ cell survival and the efficiency of porcine germ cell differentiation were moderately improved ex vivo. We assume that further optimization can be obtained with concomitant modifications in culture media components.

In microfluidics, flow focusing is widely used to produce water-in-oil droplets in microchannels at high frequency. We here report an experimental study of droplet formation in a microfluidic cross-junction with a minimum number of geometrical parameters. We mostly focus on the squeezing regime, which is composed of two distinct steps: filling and pinching. The duration of each step (and corresponding volumes of each liquid phase) is analyzed. They vary according to both water and oil flow rates. These variations provide several insights about the fluid flows in both phases. We propose several scaling laws to relate the droplet volume and frequency to the flow rate of both phases. We also discuss the influence of surfactant and channel compliance on droplet formation.

We developed a microfluidic device for the detection of bio-molecules. The active part of the device is a biofunctionalized interdigitated capacitive sensor. The microsystem consists of a sensor on silicon chip, a microfluidic channel formed by photo-patternable resist and a plastic cover. We implemented a low temperature packaging process to assemble the sensor and prevent the biological material from degradation.