To sort living cells according to our needs, it is important to understand how degree cell property measured for cell sorting fluctuates in time. Mechanical property of cells is one of essential indicators for cell sorting. Thus, in this study, we attempted to measure a time evolution of viscoelastic property such as complex shear modulus, G∗ of single cells adhered on substrates using atomic force microscopy (AFM). We observed that the G∗ largely fluctuated in time even the cells are placed on substrates in a confined condition. This indicates that in mechanical cell sorting, mechanical fluctuations of cells should be carefully estimated so that cells are precisely separated by taking the measured data involving cell fluctuations into account.