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

ABSTRACT: 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.

SUBMITTER: Chu L

PROVIDER: S-EPMC7450044 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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

Chu Ling L Tyson Jonathan J Shaw Juliana E JE Rivera-Molina Felix F Koleske Anthony J AJ Schepartz Alanna A Toomre Derek K DK

Nature communications 20200826 1

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 ...[more]

PMID: 32848153

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