This work presents the development of a micro-electro-fluidic probe (MeFP) platform as an affordable and flexible microfluidic tool for the transfection of single cells via electroporation. The platform constitutes of a 3D printed MeFP -- gold-coated microfluidic probe (MFP) with an array of pin shaped microelectrodes integrated on its tip -- and an ITO coated cell culture substrate. This setup, and submicron feature size of the MeFP, allows for a selective exposure of the targeted cell to both the electric field and hydrodynamic flow confinement (HFC) of an intercalating agent, to demonstrate transmembrane molecule delivery through electroporation. Results show successful transfer of propidium iodide (PI) through the membranes of single HeLa cells with an applied DC rectangular pulse– a proof-of-concept for MeFP’s application in delivering nucleic acids into eukaryotic cells (transfection). By adjusting the size of the HFC (varying injection and aspiration flow ratio), we show that the cell target area can be dynamically increased from the single cell footprint, to cover multiple cells. Finite Element model show that even with such low applied voltages (0.5- 3Vpk-pk), the electric field generated reach the reversible electroporation threshold. These results demonstrate the MeFP as an advancement to the currently available transfection technologies for gene therapy; delivery of DNA vaccines, in vitro fertilization, cancer treatment, regenerative medicine, and induced pluripotent stem (iPS) cells.