Various studies carried out on different photo anthropometry, but each one had some deficiencies which during the years they have been resolved. The objective of this paper is to test the efficiency of two-dimensional image processing software in photo anthropometry of hand. In this applied research, 204 office workers and industrial workers were selected. Their hands were measured by manual with photo anthropometric methods. In this study, designing the “Hand Photo Anthropometry Set,” we tried to fix the angle and distance of the camera in all of the photos. Thus, some of the common mistakes in photo anthropometric method got controlled. The taken photos were analyzed by Digimizer software, version 4.1.1.0 and Digital Caliper (Model: Mitutoyo Corp., Tokyo, Japan) was used via manual method. t-test statistical test on data revealed that there is no significant difference between the manual and photo anthropometric results (P > 0.05) and the correlation coefficients for hand dimensions are similar in both methods illustrated in the range of 0.71-0.95. The statistical analyses showed that photo anthropometry can be replaced with manual methods. Furthermore, it can provide a great help to develop an anthropometric database for work gloves manufacturers. Since the hand anthropometry is a necessary input for tool design, this survey can be used to determine the percentiles of workers’ hands.