Ultrastructure Expansion Microscopy (U-ExM) of the Fission Yeast Schizosaccharomyces pombe.

Abstract

Among widely used super-resolution microscopy techniques, including stimulated emission depletion (STED), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM), expansion microscopy (ExM) is unique in achieving increased resolution through a physical manipulation of the actual sample rather than optics or postprocessing. Originally developed for applications in neuroscience, ExM now has a solid foothold across many fields and model systems, and has been adapted to work for organisms with cell walls, including budding and fission yeasts, through the inclusion of a pre-expansion enzymatic digestion step. A variant of the ExM technique optimized for preserving the architecture of protein complexes, ultrastructure expansion microscopy (U-ExM), enables super-resolution imaging of full 3D volumes at increased throughput using conventional microscopes and can be readily combined with commonly used antibodies, dyes, and stains. Here, we present its application to the fission yeast Schizosaccharomyces pombe.