Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Minsu Kang; Jooyong Lee; Sangyoon Ko; Sang Hee Shim

doi:10.1002/cbic.202000772

Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Minsu Kang
, Jooyong Lee
, Sangyoon Ko
, Sang Hee Shim^*

^*Corresponding author for this work

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

11 Citations (Scopus)

Abstract

Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ∼5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution.

Original language	English
Pages (from-to)	1396-1399
Number of pages	4
Journal	ChemBioChem
Volume	22
Issue number	8
DOIs	https://doi.org/10.1002/cbic.202000772
Publication status	Published - 2021 Apr 16

Bibliographical note

Funding Information:
We thank the Hyunwoo Rhee Lab (Seoul National University) for providing biotin ligase plasmids and the Hak Joong Kim Lab (Korea University) for providing the Avi plasmids. This work is supported by IBS-R023-D1.

Publisher Copyright:
© 2020 Wiley-VCH GmbH

Keywords

expansion microscopy
fluorescence
single-molecule localization microscopy
super-resolution microscopy

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Organic Chemistry

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10.1002/cbic.202000772

Cite this

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title = "Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error",

abstract = "Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ∼5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution.",

keywords = "expansion microscopy, fluorescence, single-molecule localization microscopy, super-resolution microscopy",

author = "Minsu Kang and Jooyong Lee and Sangyoon Ko and Shim, \{Sang Hee\}",

note = "Funding Information: We thank the Hyunwoo Rhee Lab (Seoul National University) for providing biotin ligase plasmids and the Hak Joong Kim Lab (Korea University) for providing the Avi plasmids. This work is supported by IBS-R023-D1. Publisher Copyright: {\textcopyright} 2020 Wiley-VCH GmbH",

year = "2021",

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doi = "10.1002/cbic.202000772",

language = "English",

volume = "22",

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journal = "ChemBioChem",

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AU - Kang, Minsu

AU - Lee, Jooyong

AU - Ko, Sangyoon

AU - Shim, Sang Hee

N1 - Funding Information: We thank the Hyunwoo Rhee Lab (Seoul National University) for providing biotin ligase plasmids and the Hak Joong Kim Lab (Korea University) for providing the Avi plasmids. This work is supported by IBS-R023-D1. Publisher Copyright: © 2020 Wiley-VCH GmbH

PY - 2021/4/16

Y1 - 2021/4/16

N2 - Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ∼5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution.

AB - Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) has a potential for approaching molecular resolution. However, ExSMLM faces multiple challenges such as loss of fluorophores and proteins during polymerization, digestion or denaturation, and an increase in linkage error arising from the distance between the fluorophore and the target molecule. Here, we introduce a trifunctional streptavidin to link the target, fluorophore and gel matrix via a biotinylizable peptide tag. The resultant ExSMLM images of vimentin filaments demonstrated high labeling efficiency and a minimal linkage error of ∼5 nm. Our ExSMLM provides a simple and practical means for fluorescence imaging with molecular resolution.

KW - expansion microscopy

KW - fluorescence

KW - single-molecule localization microscopy

KW - super-resolution microscopy

UR - https://www.scopus.com/pages/publications/85099803207

U2 - 10.1002/cbic.202000772

DO - 10.1002/cbic.202000772

M3 - Article

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JF - ChemBioChem

IS - 8

ER -

Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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