A hand-held fiber optic probe instrument based on a combination of fluorescence and reflectance spectroscopy is designed to be used as an intraoperative guiding tool. Multiple light sources with fast pulse modulation are employed to deliver UV/visible light to the sample to precise tumor delineation. Remitted light from the tissue includes diffuse reflectance and fluorescence and autofluorescence enable precise quantification of endogenous and exogenous fluorophores, as well as its photobleachning products. This instrument was mainly developed for brain tumor resection guidance to assist the operating surgeon with an intuitive and real time surgical guidance tool. Further improvement and studies could be made to investigate the potential of the instrument.