Ten-fold Robust Expansion Microscopy

Hugo G.J. Damstra; Boaz Mohar; Mark Eddison; Anna Akhmanova; Lukas C. Kapitein; Paul W. Tillberg

doi:10.21769/BioProtoc.4698

Ten-fold Robust Expansion Microscopy

Hugo G.J. Damstra, Boaz Mohar, Mark Eddison, Anna Akhmanova, Lukas C. Kapitein^*, Paul W. Tillberg^*

^*Corresponding author for this work

Howard Hughes Medical Institute

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Expansion microscopy (ExM) is a powerful technique to overcome the diffraction limit of light microscopy that can be applied in both tissues and cells. In ExM, samples are embedded in a swellable polymer gel to physically expand the sample and isotropically increase resolution in x, y, and z. By systematic exploration of the ExM recipe space, we developed a novel ExM method termed Ten-fold Robust Expansion Microscopy (TREx) that, as the original ExM method, requires no specialized equipment or procedures. TREx enables ten-fold expansion of both thick mouse brain tissue sections and cultured human cells, can be handled easily, and enables high-resolution subcellular imaging with a single expansion step. Furthermore, TREx can provide ultrastructural context to subcellular protein localization by combining antibody-stained samples with off-the-shelf small molecule stains for both total protein and membranes.

Original language	English
Article number	e4698
Pages (from-to)	1-8
Number of pages	8
Journal	Bio-protocol
Volume	13
Issue number	12
DOIs	https://doi.org/10.21769/BioProtoc.4698
Publication status	Published - 20 Jun 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors.

Funding

This protocol is based on the publication of Ten-fold Robust Expansion microscopy (TREx) (Damstra et al., 2022). A.A. is supported by the Netherlands Organization for Scientific Research Spinoza Prize. L.C.K. is supported by the European Research Council (ERC Consolidator Grant 819219). B.M., M.E., and P.W.T. are supported by the Howard Hughes Medical Institute (HHMI).

Funders	Funder number
Netherlands Organization for Scientific Research Spinoza Prize
Howard Hughes Medical Institute
European Research Council	819219

Keywords

Antibody labeling
Expansion microscopy
Light microscopy
Protein stain
Super resolution
Ultrastructure

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Bio-protocol4698Final published version, 615 KBLicence: CC BY

Cite this

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AU - Tillberg, Paul W.

PY - 2023/6/20

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Ten-fold Robust Expansion Microscopy

Abstract

Bibliographical note

Funding

Keywords

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