Magnify is a universal molecular anchoring strategy for expansion microscopy

Aleksandra Klimas, Brendan R. Gallagher, Piyumi Wijesekara, Sinda Fekir, Emma F. DiBernardo, Zhangyu Cheng, Donna B. Stolz, Franca Cambi, Simon C. Watkins, Steven Brody, Amjad Horani, Alison L. Barth, Christopher I. Moore, Xi Ren, Yongxin Zhao

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

Expansion microscopy enables nanoimaging with conventional microscopes by physically and isotropically magnifying preserved biological specimens embedded in a crosslinked water-swellable hydrogel. Current expansion microscopy protocols require prior treatment with reactive anchoring chemicals to link specific labels and biomolecule classes to the gel. We describe a strategy called Magnify, which uses a mechanically sturdy gel that retains nucleic acids, proteins and lipids without the need for a separate anchoring step. Magnify expands biological specimens up to 11 times and facilitates imaging of cells and tissues with effectively around 25-nm resolution using a diffraction-limited objective lens of about 280 nm on conventional optical microscopes or with around 15 nm effective resolution if combined with super-resolution optical fluctuation imaging. We demonstrate Magnify on a broad range of biological specimens, providing insight into nanoscopic subcellular structures, including synaptic proteins from mouse brain, podocyte foot processes in formalin-fixed paraffin-embedded human kidney and defects in cilia and basal bodies in drug-treated human lung organoids.

Original languageEnglish
Pages (from-to)858-869
Number of pages12
JournalNature Biotechnology
Volume41
Issue number6
DOIs
StatePublished - Jun 2023

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