Immunofluorescence-based visualization of Malpighian tubule morphogenesis in Drosophila embryos

Malpighian tubules (MTs) undergo extensive morphogenetic remodeling during Drosophila melanogaster embryogenesis, making them sensitive indicators of developmental perturbations. Here, we present a protocol to analyze embryonic MT development in Drosophila using immunofluorescence. We describe steps for embryo collection, fixation, devitellinization, and immunostaining to visualize MT morphology. We then detail procedures for mounting, imaging, and analysis. For complete details on the use and execution of this protocol, please refer to Tiwari and Tapadia.¹.