Development of a single cell electroporation method using a scanning ion conductance microscope with a theta type probe pipette

Electroporation is a process for creating aqueous pathways in the lipid bilayer of cell membrane by external electric field passing on the membrane. In general, the electric field is applied between a pair of electrodes dipped in suspensions including cultured cells and buffer saline. We have developed a novel electroporation technique for introduction of materials into a single-cell by utilizing a nanopipette of a scanning ion conductance microscope (SICM). In this research, we developed electroporation method using the SICM with a theta type pipette probe that has two apertures at the edge of the pipette. One aperture of the pipette probe was used to control the pipette-surface distance and to apply pulse voltage for electroporation. Another one was used for material ejection by local electrophoresis. By using the pipette, propidium iodide was successfully introduced into a targeted single Hela cell without influencing on other cells around.