Nanoelectroporation and controllable intracellular delivery into localized single cell with high transfection and cell viability

Abstract

Physical introduction of foreign biomolecules such as genes, proteins, DNA and RNA into living cells with high efficiency is a challenging task for biological and therapeutic research. Bulk electroporation technique, where high electric field pulses were applied to millions of cells together in-between two large electrodes, though widely employed, however is nonspecific resulting in variable efficiency with low cell viability. Here we demonstrate controllable nano-electroporation platform for HeLa cell and human Caucasian Gastric Adenocarcinoma (AGS) cell to achieve high efficient bimolecular delivery with high cell viability. Our system consists of 40nm triangular Indium Tin Oxide (ITO) metal tip with 60nm electrode gap to provide high intense electric filed into the local region of the single cell membrane. Therefore biomolecules can be delivered by much enhance electrophoresis and diffusion effects during pulsing process through a small specific nano-region of the single cell, where remaining other area of the cell membrane unaffected. This microfluidic device has great ability to offer spatial, temporal and qualitative dosage control as well as very high transfection efficiency and high cell viability.