In order to obtain multiple planes’ reconstructed images of digital holographic microscopy simultaneously, a method for multi-plane imaging in digital holographic microscopy was proposed to extend the depth of imaging space. The quadratic distorted phase factor with pre-established parameters was used to act on experimental recorded digital hologram and the reconstructed images at multiple planes would be obtained simultaneously under single reconstruction. Firstly the imaging transfer function which used the quadratic distorted phase factor was deduced according to the transfer function of Fresnel imaging system and the rules of selecting frequency domain filtering parameters were determined. Secondly the experimental recorded digital hologram was utilized to conduct frequency domain filtering in aiming to eliminate zero-order diffraction and conjugate image. Finally the quadratic distorted phase factor was used to act on the filtered digital hologram to conduct Fresnel reconstruction. Compared with other methods, it needed single reconstruction to obtain multiple planes’ in-focus images by the method and the reconstruction distance could be chosen in arbitrary, also the reconstructed images could be got without the interference of the zero-order diffraction and conjugate image.