Maize stem sculptures, produced during the 16th and 17th centuries in New Spain (today, Mexico) are a clear example of the convergence of the artistic traditions from the American indigenous populations and European influence. This typology of sculptures is not limited to the Americas, as the examples found in European countries have shown. Therefore, a detailed technological investigation is required to correctly classify them. This work presents the interdisciplinary and multianalytical investigation of a 16th-century sculpture made with a maize stem preserved in Guadalajara city, Mexico. We used a set of techniques, such as CT, SEM-EDX, μ-FTIR, and μ-Raman, to study, from a macro to a micro level, the structure, the polychromy, and the modification of the sculpture. The results showed the use of maize stems, paper, and wood in the construction of the sculpture and the use of the traditional polychromy, as well as the numerous modifications that changed its appearance considerably resulting in its misclassification. We were able to associate the statue with the Cortés workshop (Mexico City region), probably produced in the decade of 1580, and track its liturgical use and historical development through the centuries.