A gentle palette of plasma membrane dyes

• 成果类型: Article
• 署名作者: Ling, Jing; Liu, Yitong; Dumoulin, Alexandre; Sheng, Danli; Fu, Yunzhe; Liu, Shuzhang; Ding, Ling; Huang, Lulu; Xi, Peng; Tang, Hongyun; Stoeckli, Esther T.; Chen, Zhixing
• 署名单位: Peking University; Peking University; University of Zurich; University of Zurich; Westlake University; Peking University; Peking University; Peking University; Peking University; University of Zurich; Chinese Academy of Sciences; Peking University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14758
• DOI: 10.1073/pnas.2504879122
• 发表日期: 2025-07-22
• 关键词: vesicular glutamate transporters; wheat-germ-agglutinin; live-cell; fluorescent-probes; GROWTH; phototoxicity; microscopy; fluorophores; cholesterol; mechanisms

摘要

Plasma membrane (PM) stains are important organelle markers for monitoring membrane morphology and dynamics. The state-of-the-art PM stains are bright, specific, fluorogenic, and compatible with superresolution imaging. However, when recording membrane dynamics using advanced fluorescence microscopes, PM is prone to photo-dynamic damage introduced by dyes due to its phospholipid bilayer nature. Here, we introduce PK Mem dyes tailored for time-lapse fluorescence imaging. By integrating triplet-state quenchers into the MemBright dyes featuring cyanine chromophores and amphiphilic zwitterion anchors, PK Mem dyes exhibited a three-fold reduction in photo-toxicity and a more than four-fold improvement in photostability in imaging experiments compared to MemBright prototypes. These dyes enable 2D and 3D imaging of live or fixed cancer cell lines and a wide range of primary cells, at the same time pair well with various fluorescent markers. PK Mem dyes can be applied to neuronal imaging in brain slices and in vivo two-photon imaging. The gentle nature of PK Mem palette enables ultralong-term recording of cell migration, cardiomyocyte beating, spermiogenesis, and axonal growth cone dynamics, which are prohibitively challenging using traditional PM dyes. Notably, PK Mem dyes are optically compatible with STED/SIM imaging, which can handily upgrade the routine of time-lapse neuronal imaging, such as growth cone tracking and mitochondrial transportations, into nanoscopic resolutions.