Bright photoactivatable fluorophores for single-molecule imaging.

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Nature Methods. 2016 Oct 24;13(12):985-8. doi: 10.1038/nmeth.4034

Bright photoactivatable fluorophores for single-molecule imaging. Lavis LabLippincott-Schwartz LabLiu (Zhe) LabTranscription ImagingTool Translation Team (T3)

Lavis LD, Grimm JB, English BP, Choi H, Muthusamy AK, Mehl BP, Dong P, Brown TA, Lippincott-Schwartz J, Liu Z, Lionnet T

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Abstract

Small molecule fluorophores are important tools for advanced imaging experiments. The development of self-labeling protein tags such as the HaloTag and SNAP-tag has expanded the utility of chemical dyes in live-cell microscopy. We recently described a general method for improving the brightness and photostability of small, cell-permeable fluorophores, resulting in the novel azetidine-containing "Janelia Fluor" (JF) dyes. Here, we refine and extend the utility of the JF dyes by synthesizing photoactivatable derivatives that are compatible with live cell labeling strategies. These compounds retain the superior brightness of the JF dyes once activated, but their facile photoactivation also enables improved single-particle tracking and localization microscopy experiments.