Protocol for ex vivo time lapse imaging of Drosophila melanogaster cytonemes.

This protocol describes how to image time and spatially resolved time lapses of Drosophila air sac primordium (ASP) cytonemes in cultures of wing imaginal discs. It describes how to manually measure the length of cytonemes using custom-made FIJI/ImageJ tools, and to analyze data using R/R-Studios pipeline. It can also be used for studies of cell division, organelle localization, and protein trafficking as well as other cellular materials that can be fluorescently tagged and imaged with minimal phototoxicity.