Erdmann, RS;
Baguley, SW;
Richens, JH;
Wissner, RF;
Xi, Z;
Allgeyer, ES;
Zhong, S;
... Toomre, D; + view all
(2019)
Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags.
Cell Chemical Biology
, 26
(4)
584-592.e6.
10.1016/j.chembiol.2019.01.003.
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Abstract
Super-resolution microscopy requires that subcellular structures are labeled with bright and photostable fluorophores, especially for live-cell imaging. Organic fluorophores may help here as they can yield more photons—by orders of magnitude—than fluorescent proteins. To achieve molecular specificity with organic fluorophores in live cells, self-labeling proteins are often used, with HaloTags and SNAP-tags being the most common. However, how these two different tagging systems compare with each other is unclear, especially for stimulated emission depletion (STED) microscopy, which is limited to a small repertoire of fluorophores in living cells. Herein, we compare the two labeling approaches in confocal and STED imaging using various proteins and two model systems. Strikingly, we find that the fluorescent signal can be up to 9-fold higher with HaloTags than with SNAP-tags when using far-red rhodamine derivatives. This result demonstrates that the labeling strategy matters and can greatly influence the duration of super-resolution imaging.
| Type: | Article |
|---|---|
| Title: | Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.chembiol.2019.01.003 |
| Publisher version: | https://doi.org/10.1016/j.chembiol.2019.01.003 |
| Language: | English |
| Additional information: | Copyright © 2019 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
| Keywords: | self-labeling proteins, live-cell imaging, super-resolution microscopy, HaloTag, SNAP-tag, STED, fluorophores, nanoscopy, microscopy |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Clinical and Experimental Epilepsy |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10075937 |
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