Cargando…

A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging

Despite the urgent need to image living specimens for cutting-edge biological research, most existing fluorescent labeling methods suffer from either poor optical properties or complicated operations required to realize cell-permeability and specificity. In this study, we introduce a method to overc...

Descripción completa

Detalles Bibliográficos
Autores principales: Han, Yubing, Zhang, Zhimin, Liu, Wenjie, Yao, Yuanfa, Xu, Yingke, Liu, Xu, Kuang, Cuifang, Hao, Xiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7843436/
https://www.ncbi.nlm.nih.gov/pubmed/33520932
http://dx.doi.org/10.3389/fchem.2020.601436
_version_ 1783644142355611648
author Han, Yubing
Zhang, Zhimin
Liu, Wenjie
Yao, Yuanfa
Xu, Yingke
Liu, Xu
Kuang, Cuifang
Hao, Xiang
author_facet Han, Yubing
Zhang, Zhimin
Liu, Wenjie
Yao, Yuanfa
Xu, Yingke
Liu, Xu
Kuang, Cuifang
Hao, Xiang
author_sort Han, Yubing
collection PubMed
description Despite the urgent need to image living specimens for cutting-edge biological research, most existing fluorescent labeling methods suffer from either poor optical properties or complicated operations required to realize cell-permeability and specificity. In this study, we introduce a method to overcome these limits—taking advantage of the intrinsic affinity of bright and photostable fluorophores, no matter if they are supposed to be live-cell incompatible or not. Incubated with living cells and tissues in particular conditions (concentration and temperature), some Atto and BODIPY dyes show live-cell labeling capability for specific organelles without physical cell-penetration or chemical modifications. Notably, by using Atto 647N as a live-cell mitochondrial marker, we obtain 2.5-time enhancement of brightness and photostability compared with the most commonly used SiR dye in long-term imaging. Our strategy has expanded the scientist's toolbox for understanding the dynamics and interactions of subcellular structures in living specimens.
format Online
Article
Text
id pubmed-7843436
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-78434362021-01-30 A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging Han, Yubing Zhang, Zhimin Liu, Wenjie Yao, Yuanfa Xu, Yingke Liu, Xu Kuang, Cuifang Hao, Xiang Front Chem Chemistry Despite the urgent need to image living specimens for cutting-edge biological research, most existing fluorescent labeling methods suffer from either poor optical properties or complicated operations required to realize cell-permeability and specificity. In this study, we introduce a method to overcome these limits—taking advantage of the intrinsic affinity of bright and photostable fluorophores, no matter if they are supposed to be live-cell incompatible or not. Incubated with living cells and tissues in particular conditions (concentration and temperature), some Atto and BODIPY dyes show live-cell labeling capability for specific organelles without physical cell-penetration or chemical modifications. Notably, by using Atto 647N as a live-cell mitochondrial marker, we obtain 2.5-time enhancement of brightness and photostability compared with the most commonly used SiR dye in long-term imaging. Our strategy has expanded the scientist's toolbox for understanding the dynamics and interactions of subcellular structures in living specimens. Frontiers Media S.A. 2021-01-15 /pmc/articles/PMC7843436/ /pubmed/33520932 http://dx.doi.org/10.3389/fchem.2020.601436 Text en Copyright © 2021 Han, Zhang, Liu, Yao, Xu, Liu, Kuang and Hao. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Han, Yubing
Zhang, Zhimin
Liu, Wenjie
Yao, Yuanfa
Xu, Yingke
Liu, Xu
Kuang, Cuifang
Hao, Xiang
A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging
title A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging
title_full A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging
title_fullStr A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging
title_full_unstemmed A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging
title_short A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging
title_sort labeling strategy for living specimens in long-term/super-resolution fluorescence imaging
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7843436/
https://www.ncbi.nlm.nih.gov/pubmed/33520932
http://dx.doi.org/10.3389/fchem.2020.601436
work_keys_str_mv AT hanyubing alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT zhangzhimin alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT liuwenjie alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT yaoyuanfa alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT xuyingke alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT liuxu alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT kuangcuifang alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT haoxiang alabelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT hanyubing labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT zhangzhimin labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT liuwenjie labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT yaoyuanfa labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT xuyingke labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT liuxu labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT kuangcuifang labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging
AT haoxiang labelingstrategyforlivingspecimensinlongtermsuperresolutionfluorescenceimaging