Localization of Proteins in the Secretory Pathway of Pichia pastoris Using Fluorescence Microscopy

Fluorescence microscopy is a widely used technique to localize and monitor intracellular proteins. Here we present different methods established for Pichia pastoris (syn Komagataella spp.), like immunofluorescence microscopy, the use of fluorescent fusion proteins, and various dyes to stain organelles. Furthermore, the new super-resolution technique STED is presented. The described methods mainly focus on studying the secretory pathway but are in principle applicable also to other research questions.