Background: Normal and cancerous tissues have distinct auto-fluorescence properties because of differences in their biophysical and biochemical agents. Scientific evidences related to diagnostic fluorescence imaging for detection of oral precancerous and cancerous lesions are very limited. Objectives: The aim of this study was to find out potential relationships between serum, salivary and tissue protoporphyrin IX ( PX) levels in subjects with or without oral precancerous and cancerous lesions. Also, to find out diagnostic value of fluorescence imaging (VELscope® system, LED Dental Inc., White Rock, B.C.) and salivary protoporphyrin IX (PX) in oral precancerous and cancerous lesions. Furthermore this study attempts to find out diagnostic value of the combination of approaches of fluorescence imaging and salivary protoporphyrin for detection of oral precancerous and cancerous lesions. Material and Methods: The study sample comprised 3 test groups, with biopsy confirmed precancerous (leukoplakia and lichen planus) and cancerous lesions (squamous cell carcinoma) and one control group of 25 healthy individuals. To find out sensitivity and specificity, another 100 patients presenting for routine dental care were selected and clinical examinations were followed by fluorescence imaging and normal photography, which were finally confirmed by biopsy. The clinical and histopathogical examinations were done in conjunction with photography of the oral cavity using digital camera and fluorescence imaging. Serum, tissue and salivary protoporphyrin (PX) levels were measured. Results: Using fluorescence imaging, oral cancerous and precancerous lesions showed deep purple to deep brown and dark green colour respectively, while normal tissues showed pale green colour in contrast. The PX levels in serum, salivary and tissues were significantly higher in precancerous and cancerous lesions as compared to normal healthy tissues. Salivary and serum PX levels were highly correlated in all groups. The sensitivity and specificity to the discrimination of precancerous and cancerous lesions from the healthy tissues were higher by combination approaches of salivary protoporphyrin X and VELscope® system as compared individual approach. Conclusions: Combination approach of salivary protoporphyrin X and VELscope® system are more sensitive and specific to discriminate precancerous and cancerous lesions from the healthy tissues as compared to individual approach. Further studies are required on large samples of oral precancerous and cancerous lesions to test sensitivity and specificity and thus validate the clinical applicability of fluorescence imaging in (pre)cancerous diagnostics.