Biomolecular delivery into single living cell with high transfection efficiency and high cell viability is of great interest for cellular research, therapeutics and diagnostics purpose. Here we demonstrate localize single cell electroporation using Indium Tin Oxide (ITO) based nanoelectrode chip, which is a compact, easy to use, with low voltage applications. We fabricated 40 array of nanoelectrodes on top of the silicon surface by using lithographic process and the gap between two nanoelectrodes were 70 nm. Due to apply sufficient external voltage, cell membrane deform and biomolecules gently deliver into cells. The best result was achieved using Propidium iodide dye with 94% delivery efficiency and 98% cell viability for AGS cell line. © 2019 CBMS-0001.