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. Author manuscript; available in PMC: 2017 Jan 4.
Published in final edited form as: Nat Biotechnol. 2016 Jul 4;34(9):987–992. doi: 10.1038/nbt.3625

Figure 4.

Figure 4

Workflows for expansion microscopy with protein retention. Three basic sample processing workflows were explored in this paper. Top, samples are chemically fixed and stained with antibodies, using conventional immunostaining protocols, before AcX treatment at room temperature and subsequent ExM processing (gelation, proteinase K treatment, and expansion in water). Middle, samples expressing fluorescent proteins (FPs) are chemically fixed (and optionally permeabilized) before AcX treatment, and subsequent ExM processing. Bottom, samples treated with AcX, followed by gelation, are then processed with a gentle homogenization procedure (e.g., alkaline hydrolysis and denaturation, or digestion with LysC), and finally antibody staining in the expanded state.