Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
Name
s41598-021-96258-y.pdf
Description
Published version
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1.46 MB
Format
Adobe PDF
Checksum (MD5)
49bb1258b62b806e61e88665742c9fb7
Author(s)
Lee, Hsuan
Yu, Chih-Chieh
Boyden, Edward S
Zhuang, Xiaowei
Kosuri, Pallav
Date Issued
December 2021
Journal
Scientific Reports
Publisher
Springer Science and Business Media LLC
Citation
Lee, Hsuan, Yu, Chih-Chieh, Boyden, Edward S, Zhuang, Xiaowei and Kosuri, Pallav. 2021. "Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy." Scientific Reports, 11 (1).
Version
Final published version
Abstract
AbstractThe 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.
MIT Department
McGovern Institute for Brain Research at MIT
Howard Hughes Medical Institute
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
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Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/s41598-021-96258-y
