This paper presents results from the technical examination of Flower Garland with Butterfly (1652) by Michaelina Wautier (c. 1614–1689), one of only two flower paintings known to be made by her. The painting was examined through detailed visual analysis and MA-XRF scanning, complemented by microscopic analysis of a single paint cross-section. Painted only a decade after the emergence of the sub-genre of autonomous flower garlands in Antwerp, Wautier’s composition adheres to the conventions of flower garland painting while simultaneously introducing innovative motifs. This interplay of traditional and experimental approaches is also reflected in her use of materials and paint layer build-up. The results of the technical examinations contribute to our understanding of Wautier’s artistic training, revealing an influence of Antwerp-based specialized flower painters in both iconography and technique.