Oral cancer is a serious and growing problem in many developing and developed countries. To improve the cancer screening procedure, we developed a portable light-emitting-diode (LED)-induced autofluorescence (LIAF) imager that contains two wavelength LED excitation light sources and multiple filters to capture ex vivo oral tissue autofluorescence images. Compared with conventional means of oral cancer diagnosis, the LIAF imager is a handier, faster, and more highly reliable solution. The compact design with a tiny probe allows clinicians to easily observe autofluorescence images of hidden areas located in concave deep oral cavities. The ex vivo trials conducted in Taiwan present the design and prototype of the portable LIAF imager used for analyzing 31 patients with 221 measurement points. Using the normalized factor of normal tissues under the excitation source with 365 nm of the central wavelength and without the bandpass filter, the results revealed that the sensitivity was larger than 84%, the specificity was not smaller than over 76%, the accuracy was about 80%, and the area under curve of the receiver operating characteristic (ROC) was achieved at about 87%, respectively. The fact shows the LIAF spectroscopy has the possibilities of ex vivo diagnosis and noninvasive examinations for oral cancer.