Microfluidic Single-Cell Study on Arabidopsis thaliana Protoplast Fusion-New Insights on Timescales and Reversibilities.

Plant cells are omnipotent and breeding of new varieties can be achieved by protoplast fusion. Such fusions can be achieved by treatment with poly(ethylene glycol) or by applying an electric field. Microfluidic devices allow for controlled conditions and targeted manipulation of small batches of cells down to single-cell analysis. To provide controlled conditions for protoplast fusions and achieve high reproducibility, we developed and characterized a microfluidic device to reliably trap some Arabidopsis thaliana protoplasts and induced cell fusion by controlled addition of poly(ethylene glycol) (PEG, with a molecular weight of 6000). Experiments were conducted to determine the survival rate of isolated protoplasts in our microfluidic system. Afterward, PEG-induced fusion was studied. Our results indicate that the following fusion parameters had a significant impact on the fusion efficiency and duration: PEG concentration, osmolality of solution and flow velocity. A PEG concentration below 10% led to only partial fusion. The osmolality of the PEG fusion solution was found to strongly impact the fusion process; complete fusion of two source cells sufficiently took part in slightly hyper-osmotic solutions, whereas iso-osmotic solutions led to only partial fusion at a 20% PEG concentration. We observed accelerated fusion for higher fluid velocities. Until this study, it was common sense that fusion is one-directional, i.e., once two cells are fused into one cell, they stay fused. Here, we present for the first time the reversible fusion of protoplasts. Our microfluidic device paves the way to a deeper understanding of the kinetics and processes of cell fusion.

Mid-infrared GaAs/AlGaAs micro-ring resonators characterized via thermal tuning.

Micro-ring resonators with a decoupling waveguide have been manufactured from GaAs/Al0.2Ga0.8As, accommodating mid-infrared wavelengths, and were characterized via thermal tuning. A Q-factor of 1900, a thermal full width at half maximum of 8 °C, and a thermal free spectral range of 18 °C have been achieved. The low Q-factor indicates comparatively high coupling efficiency from the input waveguide into the decoupling waveguide. The micro-ring resonators shown herein are suitable structures for advanced mid-infrared chem/bio sensing strategies via resonant-cavity enhancement. In addition, they offer high spectral resolution for evanescent field sensing strategies via effective wavelength de-multiplexing waveguide structures.