Abstract

Approaching sub-cellular biological problems requires the incorporation of new technologies and readouts. Due to their low-invasiveness, nanotechnology-based tools hold great promise for single-cell manipulation. In this talk, we discuss the incorporation of electrical measurements into nanopipette technology. Here we will introduce a newly developed single-cell manipulation platform (SU-10) using a nanopipette for single-cell injection into living cells. This newly developed technology positions its nanopipette with nanoscale precision allowing injection of minute amounts of material (drugs, proteins, CRISPR Cas9) into individual cells without comprising cell viability. This strategy will help in basic researchers in drug discovery.Unlike traditional micropipette injection techniques, the method relies not on pressure, but on applied voltage to expel liquid from the nanopipette. The distance of the nanopipette from the surface is controlled by applying a constant voltage between an electrode in the pipette and another electrode in the bath. The detection of the cell surface and the penetration are based on scanning ion conductance microscopy. The integrated software controls the movement of the nanopipette, continuing to approach the cell until a drop in the ionic current is detected, indicating the tip is at close proximity to the surface of the cell. Reduction of the electric current is detected the software stops movement in the direction of the cell and lowers the nanopipette at high speed (100 _m/s). This movement inserts the nanopipette into the cell membrane. The voltage applied to the nanopipette is then switched to inject the material into the cell. The whole process is fully automated.