Background and ObjectivesParathyroid gland detection is a fundamental skill in endocrine surgery that is enhanced with experience. This study aims to investigate the impact of near‐infrared autofluorescence (NIRAF) imaging on a surgical team's ability to recognize parathyroid glands during thyroidectomy and parathyroidectomy procedures across different training levels.MethodsPatients who underwent thyroidectomy or parathyroidectomy under NIRAF guidance by three surgeons between March and June 2024 were included. Attending surgeons, endocrine surgery fellows, and general surgery residents were assessed regarding their ability to recognize parathyroid glands before and after NIRAF imaging during the procedures. Wilcoxon and Chi‐Square tests were used for statistical analyses.ResultsAssessments were made in 32 thyroidectomy and 53 parathyroidectomy procedures, with eventual intraoperative identification of a total of 255 parathyroid glands. A total of 896 collective assessments were made. There were 250 assessments involving junior trainees, 288 involving senior trainees (fellows), and 358 involving the primary surgeon. Parathyroid detection rates for junior trainees, senior trainees, and attending surgeons before versus after NIRAF imaging were 46.5% versus 94.8%, 68% versus 97%, and 80% versus 100%, respectively (p < 0.0001). For the same groups, respectively, NIRAF imaging detected 48.3%, 29%, and 20% of parathyroid glands not initially visible on conventional view (p < 0.0001) and increased the confidence for 51.7%, 71%, and 80% of parathyroid glands that were already recognized on conventional view (p < 0.0001). Overall, for junior trainees, senior trainees, and attending surgeons, NIRAF made a positive impact in the recognition of 82.6% (n = 142/172), 62% (n = 124/200), and 42% (n = 107/255) of the total number parathyroid glands, ultimately identified during the surgical procedures respectively, (p < 0.0001).ConclusionsOur results show that NIRAF imaging improved the whole surgical team's ability to recognize parathyroid glands, with the degree of impact inversely related to the level of surgical training.