Three-dimensional (3D) stereophotogrammetry has been widely used in anthropometry for both medical and esthetic purposes. However, no studies have assessed its reliability on measuring the lower eyelid by 3D imaging. This study aimed to establish a standardized 3D anthropometric protocol for lower eyelid region and validate its reliability. Fifty-eight participants (116 eyes) were recruited with mean age of 39.14 ± 11.25 years. Two sets of VECTRA 3D images were taken for each subject, and each set of images was individually measured twice by two raters. Twenty-seven landmarks were identified in the lower eyelid region, and then 19 linear, 4 curvilinear, 7 angular and 2 areal metrics were assessed for intrarater, interrater and intramethod reliability. High reliability was found in this 3D imaging-based lower eyelid anthropometry. The mean absolute difference within 2 intrarater measurement were 0.22 and 0.08 units, the technical errors measurement were 0.31 and 0.15 units, the relative errors of measurement were 0.90% and 0.31%, the relative technical errors of measurement were 1.53% and 0.64%, and the intra-group correlation coefficient was 0.99 and 0.99. The results for interrater measurement were 0.53 units, 0.59 units, 2.94%, 3.41% and 0.96, and for intramethod measurement were 0.71 units, 0.77 units, 4.12%, 4.05% and 0.95 units, respectively. 3D stereophotogrammetry is reliable for lower eyelid anthropometry. The standardized protocol can be further applied for many purposes such as lower eyelid aging evaluation, surgical related assessment and periocular rejuvenation plan. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .