Network fixation expansion microscopy.

Authors

Yixi Liu, Chi Zhang, Corban Swain, Edward S Boyden

Published in

bioRxiv : the preprint server for biology. Oct 09, 2026. Epub Oct 09, 2026.

Abstract

Expansion microscopy (ExM) physically magnifies preserved biological specimens by using a specimen-permeating hydrogel to pull biomolecules, such as proteins, apart from each other. ExM protocols exhibit a tension between the need to initially fix proteins to each other during sample preservation, to retain proteins and their nanoscale organization, and the need to separate proteins from each other isotropically, to achieve physical magnification. We here propose to address this tension by fixing proteins directly to the hydrogel network itself, which then serves as a common substrate for both initial fixation and final separation. We equip proteins with chemical handles, then form a specimen-permeating hydrogel that binds the handles, in a ∼15-minute chemical process conducted entirely at -20°C. This network fixation ExM (nfExM) procedure enables excellent epitope retention and nanoscale organization preservation, in cultured cells. nfExM may help support ultrastructural investigations of protein complexes and interactions.

PMID:
42926696
Bibliographic data and abstract were imported from PubMed on 25 Oct 2026.

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