This paper presents a hyperspectral imaging technique based on laser-induced fluorescence for non-invasive detection of tumorous tissue on mouse skin. Hyperspectral imaging sensors collect image data in a number of narrow, adjacent spectral bands. Such high-resolution measurement of spectral information reveals contiguous emission spectra at each image pixel useful for the characterization of constituent materials. The hyperspectral image data used in this study are fluorescence images of mouse skin consisting of 21 spectral bands in the visible spectrum of the wavelengths ranging from 440 nm to 640 nm. Fluorescence signal is measured with the use of laser excitation at 337 nm. An acousto-optic tunable filter (AOTF) is used to capture images at 10 nm intervals. All spectral band images are spatially registered with the reference band image at 490 nm to obtain exact pixel correspondences by compensating the spatial offsets caused by the refraction differences in AOTF at different wavelengths during the image capture procedure. The unique fluorescence spectral signatures demonstrate a good separation to differentiate malignant tumors from normal tissues for rapid detection of skin cancers without biopsy.