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A flow-through microfluidic single cell analysis and manipulation chip is designed, fabricated, and characterized. This paper presents detailed information on the chip's design, fabrication, and characterization. The novel cell-chip is capable of electrically interfacing and interacting with single cells, thus performing precise analysis and manipulation on cell membrane permeability, which is highly important for analyzing and altering biochemical state of cells. Individual cells can either be immobilized in the chip under continuous examination to test cellular responses to specific reagents, or handled in a flow-through fashion for high-throughput cell assays, such as gene transfer and viability determination.