This study aimed to compare the accuracy of three types of apex locators versus digital radiography for working length (WL) determination. This experimental study was conducted on 58 extracted maxillary premolars. The teeth were decoronated, the access cavity was prepared, and WL was determined using a #15 K-file to serve as reference. The WL was then measured by Woodpex V, Woodpex III, and Root ZX apex locators in the presence of 0.9% saline, and also on a photostimulable phosphor plate (PSP) digital radiograph taken by the parallel technique. The values were compared with the actual WL using the paired t-test (alpha = 0.05). Digital radiography, Root ZX, Woodpex V, and Woodpex III determined the WL within ±0.5 mm from the actual value in 84.48%, 100%, 89.66%, and 87.93% of the cases, respectively. Woodpex V (p = 0.039), Woodpex III (p = 0.001), and Root ZX (p = 0.001) significantly over-estimated the WL. The WL measured on digital radiographs was not significantly different from the actual WL (p = 0.213). The position of the apical foramen (central/lateral) had no significant effect on the accuracy of WL determination by different techniques (p >0.05). Within the limitations of this in vitro study, all the tested modalities showed acceptable accuracy for WL determination in maxillary premolars.