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Deuteration improves small-molecule fluorophores

By Jonathan B. Grimm, Liangqi Xie, Jason C Casler, Ronak Patel, Ariana N. Tkachuk, Heejun Choi, Jennifer Lippincott-Schwartz, Timothy A. Brown, Benjamin S Glick, Zhe Liu, Luke Lavis

Posted 17 Aug 2020
bioRxiv DOI: 10.1101/2020.08.17.250027

Fluorescence microscopy relies on dyes that absorb short-wavelength photons and emit longer-wavelength light. In addition to this fluorescence process, dyes can undergo other photochemical reactions that result in spectral shifts and irreversible photobleaching. Increases in brightness, 'chromostability', and photostability of fluorescent dyes are therefore crucial for advancing the frontier of bioimaging. Here, we describe a general approach to improve small-molecule fluorophores using deuteration. Incorporating deuterium into the alkylamino substituents of rhodamines and other dyes improves fluorescence quantum yield, inhibits photochemically induced spectral shifts, and slows irreparable photobleaching. These compounds are easily synthesized and show improved performance in cellular imaging experiments. ### Competing Interest Statement The authors declare the following competing financial interest: Patents and patent applications describing azetidine- and deuterium-containing fluorophores (with inventors J.B.G. and L.D.L.) are assigned to HHMI.

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