The aim of the present study was to evaluate the tactile detection threshold (TDT), the filament-prick pain detection threshold (FPT), the pressure pain threshold (PPT), and the pressure pain tolerance detection threshold (PTOL) at multiple measuring points in the orofacial region of normal subjects. Sixteen males and 16 females (age range from 20 to 41 years) participated. The TDT and the FPT were measured on the cheek skin overlying the central part of the masseter muscles (MM), on the maxillary gingiva, and at the tip of the tongue, using Semmes-Weinstein monofilaments. The PPT and PTOL were measured at the central part of the MM, using a pressure algometer. The pain intensity during the FPT, PPT and the PTOL measurements was assessed on a numeric rating scale (NRS). The tongue tip had the lowest value in TDT and FPT compared to the other sites. Females showed a significantly lower TDT and FPT at the cheek skin than males. Further, measurements of PPT and PTOL confirmed the previously reported higher thresholds in males. In contrast, while the intra-oral threshold measurements revealed no gender differences, a significantly higher pain perception as evaluated using NRS, was seen in the males. A strong correlation was found between the pain responses at the same measuring site (FPT, PPT, and PTOL over the MM). In addition, the TDT and the pain responses were also correlated positively.