By the acousto-optic tunable filter (AOTF), the wavelength of the optical signal can be tuned fast in the wide spectral area. In the past study, the performance of the diffraction efficiency, the spectral resolution and the image stabilization is limit. It restricts the development of the AOTF on the spectral imaging area. In this study, we introduce the self-design non-collinear AOTF as the spetroscopic element, and set up the spectral imaging system. Further more, we obtain the microscopic images and the corresponding spectra of the plam skin tissue. In the whole working waveband, the image definition and stabilization is well,and the spectral resolution keeps on the higher level. This study indicates the applicability of the AOTF on the spectral imaging. It provides a new technology for the development of the biomedicine microscopic spectral imaging.