3D printed Micro-Electro-Fluidic Probe (MeFP) for single cell electroporation

摘要

This work presents the development of a micro-electro-fluidic probe (MeFP) platform as an affordable and flexiblemicrofluidic 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 ITOcoated cell culture substrate. This setup, and submicron feature size of the MeFP, allows for a selective exposure of thetargeted cell to both the electric field and hydrodynamic flow confinement (HFC) of an intercalating agent, todemonstrate transmembrane molecule delivery through electroporation. Results show successful transfer of propidiumiodide (PI) through the membranes of single HeLa cells with an applied DC rectangular pulse– a proof-of-concept forMeFP’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 thesingle 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 MeFPas an advancement to the currently available transfection technologies for gene therapy; delivery of DNA vaccines, invitro fertilization, cancer treatment, regenerative medicine, and induced pluripotent stem (iPS) cells.