The purpose of this study was to evaluate the effect of labial and palatinal gingiva on laser Doppler pulpal blood flow measurements. Twenty upper central incisor teeth determined to be vital from 20 volunteers were included into the study. To stabilize the probe, a laboratory-made acrylic splint positioned the probe labially on the cervical third of the tooth crown. We collected measurements without any gingival covering, and under three conditions: after application of an opaque paste on the labial gingiva, the palatal gingiva, or both the labial and palatal gingiva. The measurement values decreased 46% when there was opaque paste on labial gingiva, 10% on palatinal gingiva, and 63% on both. The decrease in the values when there was an opaque paste on palatinal gingiva was not statistically significant (p > 0.05). The differences between all other groups were statistically significant (p < 0.05). In these measurements, the contribution of labial gingiva to laser Doppler pulpal blood flow measurements is more than that of palatinal gingiva.