A technique about non-axial-scanning microscopic cell depth imaging is proposed. Fresnel lens are added into the microscopic system, and the chromatic aberration of a Fresnel lens is exploited to focus different excitation light wavelengths into different axial positions. It can realize the imaging of two or more focal planes. Based on the excitation light wavelengths of 405 nm and 532 nm, two imaging detectors are added into the excitation path of a conventional fluorescence microscope to detect the imaging information of the image plane corresponding to two different focal planes. A fluorescence microscopic depth imaging system with a detection depth of about 12 μm is established. The imaging depth and axial resolution of the proposed system are compared with those of a fluorescence microscopic depth imaging system based on a microscope-less Fresnel lens. Experimental results show that the addition of Fresnel lens enables simultaneous imaging of different focal planes. For the same fluorescence band, the lateral resolution of the system is ensured while the imaging depth of field is expanded, the system can achieve multi-band simultaneous detection at different depths of fluorescent biological cells.