Fluorogenic probes for live-cell imaging of the cytoskeleton

• Published in: Nature methods Vol. 11; no. 7; pp. 731 - 733
• Main Authors: Lukinavičius, Gražvydas, Reymond, Luc, D'Este, Elisa, Masharina, Anastasiya, Göttfert, Fabian, Ta, Haisen, Güther, Angelika, Fournier, Mathias, Rizzo, Stefano, Waldmann, Herbert, Blaukopf, Claudia, Sommer, Christoph, Gerlich, Daniel W, Arndt, Hans-Dieter, Hell, Stefan W, Johnsson, Kai
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.07.2014; Nature Publishing Group
• Subjects: 14; 14/34; 14/63; 631/1647/1888/1493; 631/1647/245/2225; 631/1647/328/2238; 631/92/96; Actins - chemistry; Animals; Axons - chemistry; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical Engineering/Biotechnology; brief-communication; Cell physiology; Cells; Cells, Cultured; Cytoskeleton; Cytoskeleton - ultrastructure; Cytotoxicity; Emissions; Erythrocytes - ultrastructure; Female; Fluorescence; Fluorescence microscopy; Fluorescent Dyes; Genetic aspects; HeLa Cells; Humans; Life Sciences; Male; Mice; Microscopy; Microscopy, Confocal - methods; Neurons - cytology; Probes; Proteomics; Rats; Rhodamines - chemistry; Scientific imaging; Silicon - chemistry; Tubulin - chemistry
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.2972

Far-red fluorogenic probes for live-cell imaging of either actin or tubulin are described and used for super-resolution microscopy of various structures in a variety of cell types. We introduce far-red, fluorogenic probes that combine minimal cytotoxicity with excellent brightness and photostability for fluorescence imaging of actin and tubulin in living cells. Applied in stimulated emission depletion (STED) microscopy, they reveal the ninefold symmetry of the centrosome and the spatial organization of actin in the axon of cultured rat neurons with a resolution unprecedented for imaging cytoskeletal structures in living cells.