Estimates of body composition have been derived using 3-dimensional optical imaging (3DO), but no equations to date have been calibrated using a 4-component (4C) model criterion. This investigation reports the development of a novel body fat prediction formula using anthropometric data from 3DO imaging and a 4C model. Anthropometric characteristics and body composition of 179 participants were measured via 3DO (Size Stream® SS20) and a 4C model. Machine learning was used to identify significant anthropometric predictors of body fat (BF%), and stepwise/lasso regression analyses were employed to develop new 3DO-derived BF% prediction equations. The combined equation was externally cross-validated using paired 3DO and DXA assessments (n = 158), producing a R2 value of 0.78 and a constant error of (X ± SD) 0.8 ± 4.5%. 3DO BF% estimates demonstrated equivalence with DXA based on equivalence testing with no proportional bias in the Bland–Altman analysis. Machine learning methods may hold potential for enhancing 3DO-derived BF% estimates.