The aim of this study was to assess the applicability of gray scale inversion in performing digital linear endodontic measurements. Standardized digital images were taken of 40 extracted human permanent single-rooted teeth with K-files size 15 placed at different lengths in the root canal, using the CDR Wireless ® digital system. All images were treated with Positive, a digital tool that inverts the grayscale value. Eight radiologists measured the distance between the lower limit of the rubber stop and the tip of the file on both original and positive images. After 15 days, they repeated this procedure in 50% of the image samples. The intra-and inter-examiner agreement was analyzed by intraclass correlation test, and the comparison between the two image groups and the actual lengths by the Student's t-test (α=0.05). Intra-and inter-examiner evaluation showed high reproducibility (r>0.95) of both original and positive images. No statistically significant differences (p>0.05) were found between measurements performed on original and positive images, but both differed significantly from the actual lengths (p<0.05). Gray scale inversion has low applicability in measuring endodontic files, since it did not improve the accuracy of measurements.