PUBLICATION
A highly stable monomeric red fluorescent protein for advanced microscopy
- Authors
- Xiong, H., Chang, Q., Ding, J., Wang, S., Zhang, W., Li, Y., Wu, Y., Lin, P., Yang, C., Liu, M., Fang, G., Yang, Y., Xie, J., Qi, D., Jiang, T., Fu, W., Hu, F., Chen, Y., Yue, R., Li, Y., Cui, Y., Li, M., Fan, S., Yang, Y., Xu, Y., Li, D., Zhang, F., Zhao, H., Wu, C., Zheng, Q., Piatkevich, K.D., Fu, Z.
- ID
- ZDB-PUB-250418-18
- Date
- 2025
- Source
- Nature Methods : (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
- none
- PubMed
- 40247125 Full text @ Nat. Methods
Citation
Xiong, H., Chang, Q., Ding, J., Wang, S., Zhang, W., Li, Y., Wu, Y., Lin, P., Yang, C., Liu, M., Fang, G., Yang, Y., Xie, J., Qi, D., Jiang, T., Fu, W., Hu, F., Chen, Y., Yue, R., Li, Y., Cui, Y., Li, M., Fan, S., Yang, Y., Xu, Y., Li, D., Zhang, F., Zhao, H., Wu, C., Zheng, Q., Piatkevich, K.D., Fu, Z. (2025) A highly stable monomeric red fluorescent protein for advanced microscopy. Nature Methods. :. Epub ahead of print.
Abstract
The stability of fluorescent proteins (FPs) is crucial for imaging techniques such as live-cell imaging, super-resolution microscopy and correlative light and electron microscopy. Although stable green and yellow FPs are available, stable monomeric red FPs (RFPs) remain limited. Here we develop an extremely stable monomeric RFP named mScarlet3-H and determine its structure at a 1.5 Å resolution. mScarlet3-H exhibits remarkable resistance to high temperature, chaotropic conditions and oxidative environments, enabling efficient correlative light and electron microscopy imaging and rapid (less than 1 day) whole-organ tissue clearing. In addition, its high photostability allows long-term three-dimensional structured illumination microscopy imaging of mitochondrial dynamics with minimal photobleaching. It also facilitates dual-color live-cell stimulated emission depletion imaging with a high signal-to-noise ratio and strong specificity. Systematic benchmarking against high-performing RFPs established mScarlet3-H as a highly stable RFP for multimodality microscopy in cell cultures and model organisms, complementing green FPs for multiplexed imaging in zebrafish, mice and Nicotiana benthamiana.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping