Single-cell electroporation.

摘要: Using modified patch-clamp methodology, we demonstrated that it is possible to insert genes or other compounds routinely into single cells by electroporation. When the cell indented a small-tipped microelectrode, voltage of 10 V less in pipette divided resistance and series cleft between tip surface. The at membrane can be high enough cause localized dielectric breakdown create pores allow enter cell. Rectangular pulses from 20 µs more than 300 ms are effective, as frequencies DC 5 kHz. most significant parameter was total time for which applied. Pipette voltages 2–10 were required, with larger requiring voltages. With optimal parameters, transfection rates excess 80% also routinely. This approach offers an effective alternative intracellular pressure injection iontophoresis placing genes, drugs, cells. Because small size electrode tips, substances inserted almost any location on their surfaces. In addition, tips electroporated only limited area so did little damage.