Measurements of the root canal during endodontic treatment have a significant influence on the course of the therapeutic process as well as on its final result in both human and veterinary medicine. The apical constriction should be the termination point for the preparation and filling of the root canal. This research was conducted with the use of a Septodont kit consisting of a small chamber filled with the examined solution in which a healthy second incisor was placed. The step back method was applied for the root canal preparation and master apical file of 30 was used. The working length was 22 mm. The examination was conducted with the use of steel as well as nickel titanium hand instruments. Different irrigation solutions and two types of apex locators were used. Measurements of the working length of the root canal showed dependence on the size of the instrument. Examinations carried out in various environments showed that analogical measurements were obtained only for sodium hypochlorite solutions. In other environments the measured sections were shortened. Comparative examinations with the use of steel instruments demonstrated insignificant measurement differences. Compared to these results, the measurements in nickel titanium group were characterized by more considerable deviations.