Journal Article Ppublish 6 citations
PubMed: 40663602 DOI: 10.1073/pnas.2504879122
A gentle palette of plasma membrane dyes.
Proc Natl Acad Sci U S A 122(29):e2504879122
Authors: Jing Ling, Yitong Liu, Alexandre Dumoulin, Danli Sheng, Yunzhe Fu, Shuzhang Liu, Ling Ding, Lulu Huang, Peng Xi, Hongyun Tang, Esther T Stoeckli, Zhixing Chen
Affiliations
- Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China. College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China.
- Department of Molecular Life Sciences, University of Zurich, Zurich 8057, Switzerland.
- Neuroscience Center Zurich, University of Zurich, Zurich 8057, Switzerland.
- School of Life Sciences, Westlake University, Hangzhou 310030, China.
- Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China.
- College of Chemistry and Molecular Engineering, Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, PKU-IDG/McGovern Institute for Brain Research, Peking University, Beijing 100871, China.
- Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, China.
- Peking University-Tsinghua University-National Institute of Biological Sciences Joint Graduate Program, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.
- University Research Priority Program "Adaptive Brain Circuits in Development and Learning", University of Zurich, Zurich 8057, Switzerland.
- Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China. College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China. PKU-Nanjing Institute of Translational Medicine, Nanjing 211800, China. Genvivo Biotech, Nanjing 211800, China. Beijing National Laboratory for Molecular Sciences, Beijing 100190, China. State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, China.
Abstract
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 photodynamic 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 phototoxicity 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.
MeSH
Cells, Growth, Growth Cones, Cell Membrane, Membranes, Plasma