Format

Send to

Choose Destination
Angew Chem Int Ed Engl. 2017 Aug 21;56(35):10408-10412. doi: 10.1002/anie.201704783. Epub 2017 Jul 24.

Long-Term Live-Cell STED Nanoscopy of Primary and Cultured Cells with the Plasma Membrane HIDE Probe DiI-SiR.

Author information

1
Department of Chemistry, Yale University, 225 Prospect Street, New Haven, CT, 06511, USA.
2
Department of Molecular Biophysics and Biochemistry and Interdepartmental Neuroscience Program, Yale University, 333 Cedar Street, New Haven, CT, 06511, USA.
3
Department of Cell Biology, Yale University, 333 Cedar Street, New Haven, CT, 06511, USA.

Abstract

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.

KEYWORDS:

bioorthogonal chemistry; fluorophores; membranes; neurons; super-resolution microscopy

PMID:
28679029
PMCID:
PMC5576494
DOI:
10.1002/anie.201704783
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Wiley Icon for PubMed Central
Loading ...
Support Center