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Impedance analysis of single cells offers the possibility of obtaining accurate, reliable, and in-depth information about their pathological condition. In this work, we present the physical capture of a single cell using a microfluidic device with a three-pillar microstructure, impedance measurement using a set of planar microelectrodes, and the derivation of an electrical model that fits the experimental results. The relationship between impedance characteristics and the location of a single HeLa cell (human cervical epithelioid carcinoma) is investigated by impedance spectroscopy. When a single cell fell toward the electrodes after it has been trapped by the three-pillar microstructure, the impedance and the phase change are measured in an operating frequency range of 1 to 100 kHz. The equivalent circuit model is established and two elements that depend on cell location are used to investigate the impedance change of a single HeLa cell.