Hollow nanoneedle array and its utilization for repeated administration of biomolecules to the same cells

We present a novel hollow nanoneedle array (NNA) device capable of simultaneously delivering diverse cargo into a group of cells in a culture over prolonged periods. The silica needles are fed by a common reservoir whose content can be replenished and modified in real time while maintaining contact with the same cells. The NNA, albeit its submicrometer features, is fabricated in a silicon-on-insulator wafer using conventional, large scale, silicon technology. 3T3-NIH fibroblast cells and HEK293 human embryonic kidney cells are shown to grow and proliferate successfully on the NNAs. Cargo delivery from the reservoir through the needles to a group of HEK293 cells in the culture is demonstrated by repeated administration of fluorescently labeled dextran to the same cells and transfection with DNA coding for red fluorescent protein. The capabilities demonstrated by the NNA device open the door to large scale studies of the effect of selected cells on their environment as encountered, for instance, in the study of cell-fate decisions, the role of cell-autonomous versus nonautonomous mechanisms in developmental biology, and in the study of excitable cell-networks.