To determine the extensions of cavities prepared conventionally by bur or by a fluorescence-controlled Er:YAG laser. Sixty-five human teeth with dentine caries were bisected through the caries lesion and were treated by a fluorescence-controlled Er:YAG laser in a non-contact or a contact mode or by a rotary bur. The specimens were subjected to histological staining and a quantitative evaluation of cavity area (mm(2)) by computer-assisted alignment. Data were tested for statistical significant differences by the Wilcoxon test (p < 0.05). Twenty-three out of 29 cavities were smaller after caries removal with the non-contact laser compared to the bur. For a threshold level of seven, a cavity size difference of 1.63 (1.86) mm(2) was calculated compared to a cavity size difference of 5.35 (5.05) mm(2) after bur excavation. The differences were statistically significant (p = 0.029). No significant differences were observed between the cavity size differences after excavation with the non-contact or the contact laser handpiece. Residual bacteria within the cavity floor were found only in low numbers after all treatments. The present in vitro study indicates that caries removal by a fluorescence-controlled Er:YAG laser using a threshold level of seven resulted in less dentine loss than preparations by a bur.