In diabetes patients and animal models, major cell morphology changes in renal tubules have been observed. The relationship between insulin and the microvilliar structure of tubular epithelial cells, however, remains obscure. We used the Scanning Ion Conductance Microscope to image microvillars in the living cell to consider microvillar dynamic. Here, on the tubular epithelial cell surface, we found two layers of microvilli: short compact microvilli, and net-like long microvilli. The length and density of microvilli may be improved by insulin treatment .The PI3K/PLCγ signaling pathway, rather than the PI3K/Arp2/3 signal pathway, mediated this operation. In conclusion, our findings present a novel transduction mechanism for insulin signaling, which contributes to the understanding of complex regulation of renal tubular epithelial cell microvilli.