Single-cell membrane surgery by virtual electrodes using an inverted electron beam lithography system

Single cell manipulation had been applied to various analyses of cell functions, however, the nanometer size surgery tools for the single cell analysis was limited the resolution and rapidly. We proposed here a single cell manipulation using a fine focused electron beam. Since the electron beam could provide virtual electrodes which promoted electric influences and they were focused in sub-micrometers, the fine manipulation could performed subcellular surgery of the single living cell. By using our method with an inverted electron beam lithography system, we reported here the electrokinetic induced poration of localized single cells and subcellular detachment. Observing this poration, the electron beam irradiation induced PI dye inflow into the single living cells and their fluorescence were confirmed. Although the viability of the treated cells was still unclear, the poration probability by our method was demonstrated.