Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 22, Issue 21, Pages -Publisher
MDPI
DOI: 10.3390/ijms222111799
Keywords
super-resolution microscopy; PAINT; fluorescent labeling; exchangeable labels
Funding
- Ministry of Science and Higher Education of the Russian Federation [075-15-2020-773]
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Fluorescent labeling is a well-established method for visualizing cellular structures and dynamics, with super-resolution microscopy bypassing the diffraction limit in image resolution. Transiently interacting labels can provide high photostability through constant supply, while also interfering less with the intrinsic dynamics and cellular functions of labeled molecules.
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.
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