Two-color nanoscopy of organelles for extended times with HIDE probes.

Performing multi-color nanoscopy for extended times is challenging due to the rapid photobleaching rate of most fluorophores. Here we describe a new fluorophore (Yale-595) and a bio-orthogonal labeling strategy that enables two-color super-resolution (STED) and 3D confocal imaging of two organelles simultaneously for extended times using high-density environmentally sensitive (HIDE) probes. Because HIDE probes are small, cell-permeant molecules, they can visualize dual organelle dynamics in hard-to-transfect cell lines by super-resolution for over an order of magnitude longer than with tagged proteins. The extended time domain possible using these tools reveals dynamic nanoscale targeting between different organelles.

Citation

Ling Chu, Jonathan Tyson, Juliana E Shaw, Felix Rivera-Molina, Anthony J Koleske, Alanna Schepartz, Derek K Toomre. Two-color nanoscopy of organelles for extended times with HIDE probes. Nature communications. 2020 Aug 26;11(1):4271

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PMID: 32848153

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