Premium
Long‐Term Live‐Cell STED Nanoscopy of Primary and Cultured Cells with the Plasma Membrane HIDE Probe DiI‐SiR
Author(s) -
Thompson Alexander D.,
Omar Mitchell H.,
RiveraMolina Felix,
Xi Zhiqun,
Koleske Anthony J.,
Toomre Derek K.,
Schepartz Alanna
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201704783
Subject(s) - sted microscopy , temporal resolution , fluorophore , chemistry , live cell imaging , superresolution , biophysics , filopodia , fluorescence , stimulated emission , cell , optics , physics , biology , laser , biochemistry , artificial intelligence , computer science , image (mathematics)
Super‐resolution imaging of live cells over extended time periods with high temporal resolution requires high‐density labeling and extraordinary fluorophore photostability. Herein, we achieve this goal by combining the attributes of the high‐density plasma membrane probe DiI‐TCO and the photostable STED dye SiR‐Tz. These components undergo rapid tetrazine ligation within the plasma membrane to generate the HIDE probe DiI‐SiR. Using DiI‐SiR, we visualized filopodia dynamics in HeLa cells over 25 min at 0.5 s temporal resolution, and visualized dynamic contact‐mediated repulsion events in primary mouse hippocampal neurons over 9 min at 2 s temporal resolution. HIDE probes such as DiI‐SiR are non‐toxic and do not require transfection, and their apparent photostability significantly improves the ability to monitor dynamic processes in live cells at super‐resolution over biologically relevant timescales.