Protein and lipid expansion microscopy with trypsin and tyramide signal amplification for 3D imaging.

ABSTRACT: Expansion microscopy, whereby the relative positions of biomolecules are physically increased via hydrogel expansion, can be used to reveal ultrafine structures of cells under a conventional microscope. Despite its utility for achieving super-resolution imaging, expansion microscopy suffers a major drawback, namely reduced fluorescence signals caused by excessive proteolysis and swelling effects. This caveat results in a lower photon budget and disfavors fluorescence imaging over a large field of view that can cover an entire expanded cell, especially in 3D. In addition, the complex procedures and specialized reagents of expansion microscopy hinder its popularization. Here, we modify expansion microscopy by deploying trypsin digestion to reduce protein loss and tyramide signal amplificatio

SUBMITTER: Wang UT 

PROVIDER: S-EPMC10713663 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature