Objective: To measure directly the thickness of the palatal mucosa using sharp cannulas and to evaluate potential correlations with gender and body mass index (BMI). Method and Materials: Thirty-three participants (19 women) 23 to 41 years of age (27.8 ± 4.1) were examined. Mucosal thickness from maxillary left canine to left second molar was measured at defined sites under local anesthesia using singleuse cannulas with silicone stops. The cannulas were evaluated on video images. Results: The mean thickness of the palatal mucosa was lowest in the first molar areas (4.39 ± 1.05 mm), increased considerably in the canine/premolar regions, and was highest in the second molar areas (5.75 ± 1.78 mm). Women exhibited higher mucosal thickness values than men in all tooth regions (P .05 for the canine and the molar regions). Multivariate analysis revealed a relation of palatal mucosal thickness to gender and BMI. Conclusion: The thickness of the palatal mucosa is characterized by variations that correlate with tooth regions, gender, and BMI. Direct measurements with sharp cannulas provide higher thickness values than measurements with blunt periodontal probes or ultrasonographic measurements. This finding is important particularly with regard to the hard palate as a donor site for connective tissue grafts.