Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags.
Erdmann, Roman S
Baguley, Stephanie Wood
Wissner, Rebecca F
Thompson, Alexander D
Rothman, James E
Cell Chemical Biology
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Erdmann, R. S., Baguley, S. W., Richens, J., Wissner, R. F., Xi, Z., Allgeyer, E., Zhong, S., et al. (2019). Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags.. Cell Chemical Biology https://doi.org/10.1016/j.chembiol.2019.01.003
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.
HaloTag, SNAP-tag, STED, fluorophores, live-cell imaging, microscopy, nanoscopy, self-labeling proteins, super-resolution microscopy
This work was supported by a Wellcome Trust Foundation grant (095927/A/11/Z) and NIH grant (R01GM118486). R.S.E. was supported by an Advanced Postdoc Mobility Fellowship from the Swiss National Foundation.
Wellcome Trust (095927/B/11/Z)
External DOI: https://doi.org/10.1016/j.chembiol.2019.01.003
This record's URL: https://www.repository.cam.ac.uk/handle/1810/293848
All rights reserved, Attribution 4.0 International
Licence URL: http://creativecommons.org/licenses/by/4.0/
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