PT  - JOURNAL ARTICLE
AU  - Jonathan B. Grimm
AU  - Anand K. Muthusamy
AU  - Yajie Liang
AU  - Timothy A. Brown
AU  - William C. Lemon
AU  - Ronak Patel
AU  - Rongwen Lu
AU  - John J. Macklin
AU  - Phillip J. Keller
AU  - Na Ji
AU  - Luke D. Lavis
TI  - A general method to fine-tune fluorophores for live-cell and <em>in vivo</em> imaging
AID  - 10.1101/127613
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 127613
4099  - http://biorxiv.org/content/early/2017/04/14/127613.short
4100  - http://biorxiv.org/content/early/2017/04/14/127613.full
AB  - Pushing the frontier of fluorescence microscopy requires the design of enhanced fluorophores with finely tuned properties. We recently discovered that incorporation of four-membered azetidine rings into classic fluorophore structures elicits substantial increases in brightness and photostability, resulting in the ‘Janelia Fluor’ (JF) series of dyes. Here, we refine and extend this strategy, showing that incorporation of 3-substituted azetidine groups allows rational tuning of the spectral and chemical properties with unprecedented precision. This strategy yields a palette of new fluorescent and fluorogenic labels with excitation ranging from blue to the far-red with utility in live cells, tissue, and animals.