The effect of period of repeating thickness on the stress is studied in ZrO 2/SiO2 multilayers deposited by electron beam evaporation on BK7 glass a nd fused silica substrates, separately. The results show that the residual stress in the multilayers is compressive, and with the increase of the period of repeating thickness the residual stress in multilayers decrease in both BK7 and fused silica substrates. At the same time, the residual stress in multilayers deposited on BK7 glass substrates is less than that in the samples deposited on fused silica substrates. The variation of the microstructure examined by the x-ray diffraction shows that the microscopic deformation does not correspond to the macroscopic stress, which may be due to the variation of the interface stress.