Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy

Abstract The accuracy of expansion microscopy (ExM) depends on the structural preservation of samples embedded in a hydrogel. However, it has been unknown to what extent gel embedding alters the molecular positions of individual labeled sites. Here, we quantified the accuracy of gel embedding by usi...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hsuan Lee, Chih-Chieh Yu, Edward S. Boyden, Xiaowei Zhuang, Pallav Kosuri
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/07fd9716e4d444bcb23396fa05e84b9c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:07fd9716e4d444bcb23396fa05e84b9c
record_format	dspace
spelling	oai:doaj.org-article:07fd9716e4d444bcb23396fa05e84b9c2021-12-02T18:51:46ZTetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy10.1038/s41598-021-96258-y2045-2322https://doaj.org/article/07fd9716e4d444bcb23396fa05e84b9c2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96258-yhttps://doaj.org/toc/2045-2322Abstract The accuracy of expansion microscopy (ExM) depends on the structural preservation of samples embedded in a hydrogel. However, it has been unknown to what extent gel embedding alters the molecular positions of individual labeled sites. Here, we quantified the accuracy of gel embedding by using stochastic optical reconstruction microscopy (STORM) to image DNA origami with well-defined structures. We found that embedding in hydrogels based on polyacrylamide, the most widely used chemistry in ExM, resulted in random displacements of labeled sites with a standard deviation of ~ 16 nm. In contrast, we found that embedding in tetra-gel, a hydrogel that does not depend on free-radical chain-growth polymerization, preserved labeled sites with a standard deviation of less than 5 nm. By combining tetra-gel ExM with STORM, we were able to resolve 11-nm structural features without the loss in accuracy seen with polyacrylamide gels. Our study thus provides direct measurements of the single-molecule distortions resulting from hydrogel embedding, and presents a way to improve super-resolution microscopy through combination with tetra-gel ExM.Hsuan LeeChih-Chieh YuEdward S. BoydenXiaowei ZhuangPallav KosuriNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hsuan Lee Chih-Chieh Yu Edward S. Boyden Xiaowei Zhuang Pallav Kosuri Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
description	Abstract The accuracy of expansion microscopy (ExM) depends on the structural preservation of samples embedded in a hydrogel. However, it has been unknown to what extent gel embedding alters the molecular positions of individual labeled sites. Here, we quantified the accuracy of gel embedding by using stochastic optical reconstruction microscopy (STORM) to image DNA origami with well-defined structures. We found that embedding in hydrogels based on polyacrylamide, the most widely used chemistry in ExM, resulted in random displacements of labeled sites with a standard deviation of ~ 16 nm. In contrast, we found that embedding in tetra-gel, a hydrogel that does not depend on free-radical chain-growth polymerization, preserved labeled sites with a standard deviation of less than 5 nm. By combining tetra-gel ExM with STORM, we were able to resolve 11-nm structural features without the loss in accuracy seen with polyacrylamide gels. Our study thus provides direct measurements of the single-molecule distortions resulting from hydrogel embedding, and presents a way to improve super-resolution microscopy through combination with tetra-gel ExM.
format	article
author	Hsuan Lee Chih-Chieh Yu Edward S. Boyden Xiaowei Zhuang Pallav Kosuri
author_facet	Hsuan Lee Chih-Chieh Yu Edward S. Boyden Xiaowei Zhuang Pallav Kosuri
author_sort	Hsuan Lee
title	Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
title_short	Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
title_full	Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
title_fullStr	Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
title_full_unstemmed	Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
title_sort	tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/07fd9716e4d444bcb23396fa05e84b9c
work_keys_str_mv	AT hsuanlee tetragelenablessuperioraccuracyincombinedsuperresolutionimagingandexpansionmicroscopy AT chihchiehyu tetragelenablessuperioraccuracyincombinedsuperresolutionimagingandexpansionmicroscopy AT edwardsboyden tetragelenablessuperioraccuracyincombinedsuperresolutionimagingandexpansionmicroscopy AT xiaoweizhuang tetragelenablessuperioraccuracyincombinedsuperresolutionimagingandexpansionmicroscopy AT pallavkosuri tetragelenablessuperioraccuracyincombinedsuperresolutionimagingandexpansionmicroscopy
_version_	1718377368277483520

Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy

Ejemplares similares