Transient Fluorescence Labeling: Low Affinity—High Benefits

Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (ST...

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Autores principales:	Maxim M. Perfilov, Alexey S. Gavrikov, Konstantin A. Lukyanov, Alexander S. Mishin
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	super-resolution microscopy PAINT fluorescent labeling exchangeable labels Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/c23debb0f7f54e0babac0704da2cc759
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spelling	oai:doaj.org-article:c23debb0f7f54e0babac0704da2cc7592021-11-11T17:14:41ZTransient Fluorescence Labeling: Low Affinity—High Benefits10.3390/ijms2221117991422-00671661-6596https://doaj.org/article/c23debb0f7f54e0babac0704da2cc7592021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11799https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (STED) microscopy impose tight restrictions on the physico-chemical properties of labels. One of them—the requirement for high photostability—can be satisfied by transiently interacting labels: a constant supply of transient labels from a medium replenishes the loss in the signal caused by photobleaching. Moreover, exchangeable tags are less likely to hinder the intrinsic dynamics and cellular functions of labeled molecules. Low-affinity labels may be used both for fixed and living cells in a range of nanoscopy modalities. Nevertheless, the design of optimal labeling and imaging protocols with these novel tags remains tricky. In this review, we highlight the pros and cons of a wide variety of transiently interacting labels. We further discuss the state of the art and future perspectives of low-affinity labeling methods.Maxim M. PerfilovAlexey S. GavrikovKonstantin A. LukyanovAlexander S. MishinMDPI AGarticlesuper-resolution microscopyPAINTfluorescent labelingexchangeable labelsBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11799, p 11799 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	super-resolution microscopy PAINT fluorescent labeling exchangeable labels Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	super-resolution microscopy PAINT fluorescent labeling exchangeable labels Biology (General) QH301-705.5 Chemistry QD1-999 Maxim M. Perfilov Alexey S. Gavrikov Konstantin A. Lukyanov Alexander S. Mishin Transient Fluorescence Labeling: Low Affinity—High Benefits
description	Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (STED) microscopy impose tight restrictions on the physico-chemical properties of labels. One of them—the requirement for high photostability—can be satisfied by transiently interacting labels: a constant supply of transient labels from a medium replenishes the loss in the signal caused by photobleaching. Moreover, exchangeable tags are less likely to hinder the intrinsic dynamics and cellular functions of labeled molecules. Low-affinity labels may be used both for fixed and living cells in a range of nanoscopy modalities. Nevertheless, the design of optimal labeling and imaging protocols with these novel tags remains tricky. In this review, we highlight the pros and cons of a wide variety of transiently interacting labels. We further discuss the state of the art and future perspectives of low-affinity labeling methods.
format	article
author	Maxim M. Perfilov Alexey S. Gavrikov Konstantin A. Lukyanov Alexander S. Mishin
author_facet	Maxim M. Perfilov Alexey S. Gavrikov Konstantin A. Lukyanov Alexander S. Mishin
author_sort	Maxim M. Perfilov
title	Transient Fluorescence Labeling: Low Affinity—High Benefits
title_short	Transient Fluorescence Labeling: Low Affinity—High Benefits
title_full	Transient Fluorescence Labeling: Low Affinity—High Benefits
title_fullStr	Transient Fluorescence Labeling: Low Affinity—High Benefits
title_full_unstemmed	Transient Fluorescence Labeling: Low Affinity—High Benefits
title_sort	transient fluorescence labeling: low affinity—high benefits
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/c23debb0f7f54e0babac0704da2cc759
work_keys_str_mv	AT maximmperfilov transientfluorescencelabelinglowaffinityhighbenefits AT alexeysgavrikov transientfluorescencelabelinglowaffinityhighbenefits AT konstantinalukyanov transientfluorescencelabelinglowaffinityhighbenefits AT alexandersmishin transientfluorescencelabelinglowaffinityhighbenefits
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Transient Fluorescence Labeling: Low Affinity—High Benefits

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