Imaging Macrophage Fascin Bundles

UAS transgenic constructs encoding fluorescent proteins were expressed specifically in macrophages using the singed-Gal4 driver line (Zanet et al., 2012 (link)). Live embryos were mounted as previously described (Wood et al., 2006 (link)). Briefly, stage 15 embryos were dechorionated in bleach and mounted under coverslips on hydrophobic Lumox dishes (Sarstedt, Germany) in Voltalef oil 10S. Embryos were then imaged with a spinning disk microscope (UltraVIEW VoX PerkinElmer) with a 63× N.A. 1.4 objective acquiring one image every 30 s. Measurements of the length of fascin bundles were performed using Volocity software. MT arms were defined as polarised bundles of MT that anticipate direction of migration as previously described previously (Stramer et al., 2010 (link)).