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The aim of this study was to compare the quality of apical enlargement of mesiobuccal canals of mandibular molars using conventional stainless steel hand files (K files) and nickel-titanium (Ni-Ti) rotary instruments (LightSpeed). Thirty freshly extracted mandibular molars were randomly assigned to three equal groups (n = 10 each group). The mesiobuccal canals were instrumented with K file using step-back technique without coronal flaring (control; group 1), K file using step-back technique after coronal flaring (group 2), and LightSpeed instrumentation (group 3). Specific criteria for apical enlargement based on initial apical size were used. For step-back techniques, the master apical file sizes were based on the Grossman criterion of three sizes larger than the first file that bound at working length, without coronal flaring (group 1) and with flaring (group 2). For the LightSpeed (LS) group, the master apical rotary sizes were based on the manufacturer's recommendation. Canal cleanliness, canal transportation, and final canal shapes were determined histologically at 1-mm and 3-mm levels short of the working length. Canals were prepared to significantly larger sizes using LS instrumentation than with either hand instrumentation techniques (15–17 ISO units, p < 0.001). LS instrumentation allowed greater apical enlargement with significantly cleaner canals, less apical transportation, and better canal shape than both hand instrumentation groups at both levels (p < 0.05). None of the three techniques was totally effective in cleaning the apical canal space. It was concluded that greater apical enlargement using LS rotary instruments is beneficial as an attempt to further debride the apical third region in mesiobuccal canals of mandibular molars. Instrument designs, alloy properties, and canal curvature are important factors that determine the feasibility of greater apical enlargement in narrow canals.