Correlative super-resolution and electron microscopy to resolve subcellular protein localization
José María Mateos
Center for Microscopy and Image Analysis, University of Zürich (Switzerland)
Methods to determine the subcellular localization of proteins and their relationship with different compartments of the cell are essential tools to understand their functions and possible interactions. Super-resolution microscopy in combination with electron microscopy provides such information. In order to achieve the best correlation of protein expression with a specific subcellular compartment the use of the same ultrathin section for light and electron microscopy is required. Among the different sectioning methods, Tokuyasu cryo-sections preserve the antigenicity of many epitopes and provide a good tissue ultrastructure. We have introduced sample collection on silicon wafers and contrasting by platinum shadowing. These steps are clear improvements in the technique in terms of ease of use, reproducibility, and time to complete experiments. We have recently demonstrated the applicability of the method to detect nuclear pores and mitochondria proteins in mouse tissue. I will present this technique as well as new data on protein expression in the developing zebrafish retina.