The aim of this study was to evaluate the efficacy of smear layer removal from teeth following root canals using lasers (Er:YAG and Nd:YAG), NaOCl, 17% EDTA, and MTAD by scanning electron microscopy (SEM). Root canals were irrigated with 3 mL of 5.25% NaOCl after each file. Teeth were randomly divided into five groups. In the final irrigation, canals were irrigated with 5.25% NaOCl (Group 1, control), 17% EDTA (Group 2), or BioPure MTAD (Group 3). Laser groups were irradiated with Er:YAG laser (1.8 W, 120 mJ, 15 Hz) (Group 4) or Nd:YAG laser (1 W, 100 mJ, 15 Hz) (Group 5). The smear layer at the apical, middle, and coronal thirds of each root canal was imaged using SEM. Smear layer removal by EDTA was the most effective in all regions. The difference between EDTA and MTAD was statistically significant in the coronal and middle (p < 0.05), but not the apical (p > 0.05), regions. The difference between the control and laser groups was not statistically significant (p > 0.05). In all regions, the difference between 17% EDTA–MTAD and the control–laser groups was statistically significant (p < 0.05). Although improvement was observed in removal of the smear layer using alternative materials and techniques, application of a combination of EDTA and NaOCl remains an effective technique.