中国科学院生物物理研究所徐平勇小组的一项最新研究报道了用于双色3D STED和延时3D SIM成像的一种高度光稳定的单体红色荧光蛋白。相关论文于2025年11月26日发表在《自然—方法学》杂志上。

在这里，课题组介绍了mScarlet3S2，这是一种RFP，其光稳定性比其前身mScarlet3提高了29倍，并且优于其他现有的RFP。使用mScarlet3 S2，该课题组在3D STED成像中实现了150多个Z堆栈，详细揭示了内质网（ER）的结构。scarlet3 S2包括非平面ER结、核包膜（NE）内陷、ER - NE接触的三维地图、不同的接触形态（点状、带状和支状）和极化ER - NE结分布。这些发现重新定义了他们对ER-NE接口的结构理解，并证明了mScarlet3S2在揭示亚细胞复杂性。

研究人员表示，高度光稳定的红色荧光蛋白（RFPs）对于双色荧光显微镜，包括超分辨率显微镜是非常宝贵的。

附：英文原文

Title: A highly photostable monomeric red fluorescent protein for dual-color 3D STED and time-lapse 3D SIM imaging

Author: Ding, Ya, He, Wenting, Wang, Kunhao, Xue, Fudong, Zheng, Ke, Zhao, Shiqun, Qi, Dong, Li, Tianyi, Xia, Peng, Pan, Leiting, Chen, Liangyi, Chen, Zhixing, Yuan, Lin, Xu, Pingyong

Issue&Volume: 2025-11-26

Abstract: Highly photostable red fluorescent proteins (RFPs) are invaluable for dual-color fluorescence microscopy, including super-resolution microscopy. Here we present mScarlet3S2, an RFP that exhibits a 29-fold improvement in photostability over its predecessor, mScarlet3, and outperforms other existing RFPs. This high photostability enables prolonged 2D and 3D imaging using both structured illumination microscopy and stimulated emission depletion microscopy. Using mScarlet3S2, we achieved over 150 Z-stacks in 3D STED imaging, revealing the architecture of the endoplasmic reticulum (ER) in detail. Key findings facilitated by mScarlet3S2 include nonplanar ER junctions, nuclear envelope (NE) invaginations, 3D maps of ER–NE contacts, diverse contact morphotypes (punctate, ribbon-like and branched) and polarized ER–NE junction distributions. These findings redefine our structural understanding of the ER–NE interface and demonstrate the value of mScarlet3S2 in revealing subcellular complexity.

DOI: 10.1038/s41592-025-02962-2

Source: https://www.nature.com/articles/s41592-025-02962-2