Scientific question: To acquire their quasi-crystalline structures, muscles need to assemble in a highly-regulated fashion at the molecular level. The tens of proteins species have to assemble very precisely, but much has to be uncovered how they do it and how they are assembled in a mature muscle. In this workshop we will decipher where some of these proteins are localized in the Drosophila flight muscle using DNA-PAINT and automated data analysis. Practical aspects to be tackled: DNA-PAINT is a very simple and elegant way to achieve very high resolution, but key steps need to be mastered to unleash the power of the method. We will discuss the sample preparation, choosing the right DNA oligos, fluorophores, coverslip preparation, extracting fluorophores localization and drift correction.