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Bright photoactivatable fluorophores for single-molecule imaging

By Jonathan B. Grimm, Brian P English, Anand K Muthusamy, Brian P Mehl, Peng Dong, Timothy A. Brown, Zhe Liu, Timothée Lionnet, Luke Lavis

Posted 29 Jul 2016
bioRxiv DOI: 10.1101/066779 (published DOI: 10.1038/nmeth.4034)

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 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 established live cell labeling strategies. These compounds retain the superior brightness of the JF dyes but their facile photoactivation enables improved single-particle tracking and localization microscopy experiments.

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