Background. Despite of having improved endodontic file designs as well as the reinforced metal alloy file structure, intracanal endodontic file separation (EFS) is still a very problematic and worrisome dental incident, which usually occurs without any visible signs of permanent deformation. Further, there have been conflicting reports regarding the clinical significance of leaving separated files within root canals. Aims. The aim of this study was to look into the current perceptions and awareness about file separation during endodontic treatment among the dental house officers (DHOs). Materials and Methods. A novel validated questionnaire comprising of 15 close-ended questions was distributed anonymously via Google Forms through email to 1100 DHOs across Pakistan. The questionnaire consisted of two parts: the first component (Section I) collected demographic data and the second component (Section II) investigated the causes of EFS during root canal treatment. Following the completion of socioeconomic information, including age and gender, the DHOs were asked to answer a few questions about the various reasons for endodontic instrument fracture. Results. A total of 800 responses were recorded, with an effective rate of 72.8%. The majority of the DHOs ( p value < 0.001) perceived that endodontic instrument fracture occurred in the posterior (61.5%) and apical third of the canal (50.5%) and in older permanent dentition (67.3%), possibly due to patient anxiety (62%). Better choice of instrument (61.15%), operators’ experience (95.3%), knowledge (87.5%), and proper root canal cleaning (91.1%) are believed to be the vital steps in reducing endodontic file separation/fracture. Furthermore, majority of them ( p value < 0.001) perceived that stainless steel was a superior alloy for filing instruments. Manual files tend to be more prone to fractures due to repeated use than rotary files. Conclusion. This study demonstrated that young DHOs had adequate knowledge and awareness regarding the potential predisposing factors and handling techniques for EFS. This study thereby provides an evaluating tool to access the insights of the current perceptions and awareness of DHOs concerning EFS.