title: DCDHF fluorophores for single-molecule imaging in cells.
abstract: There is a persistent need for small-molecule fluorescent labels optimized for single-molecule imaging in the cellular environment. Application of these labels comes with a set of strict requirements: strong absorption, efficient and stable emission, water solubility and membrane permeability, low background emission, and red-shifted absorption to avoid cell autofluorescence. We have designed and characterized several fluorophores, termed "DCDHF" fluorophores, for use in live-cell imaging based on the push-pull design: an amine donor group and a 2-dicyanomethylene-3-cyano-2,5-dihydrofuran (DCDHF) acceptor group, separated by a pi-rich conjugated network. In general, the DCDHF fluorophores are comparatively photostable, sensitive to local environment, and their chemistries and photophysics are tunable to optimize absorption wavelength, membrane affinity, and solubility. Especially valuable are fluorophores with sophisticated photophysics for applications requiring additional facets of control, such as photoactivation. For example, we have reengineered a red-emitting DCDHF fluorophore so that it is dark until photoactivated with a short burst of low-intensity violet light. This molecule and its relatives provide a new class of bright photoactivatable small-molecule fluorophores, which are needed for super-resolution imaging schemes that require active control (here turning-on) of single-molecule emission.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, California 94305, USA.
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Lord
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Conley
authors.forename: Hsiao-Lu D
authors.initials: HL
authors.lastname: Lee
authors.forename: Stefanie Y
authors.initials: SY
authors.lastname: Nishimura
authors.forename: Andrea K
authors.initials: AK
authors.lastname: Pomerantz
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Willets
authors.forename: Zhikuan
authors.initials: Z
authors.lastname: Lu
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Na
authors.initials: N
authors.lastname: Liu
authors.forename: Reichel
authors.initials: R
authors.lastname: Samuel
authors.forename: Ryan
authors.initials: R
authors.lastname: Weber
authors.forename: Alexander
authors.initials: A
authors.lastname: Semyonov
authors.forename: Meng
authors.initials: M
authors.lastname: He
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Twieg
authors.forename: W E
authors.initials: WE
authors.lastname: Moerner
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry*; D005663:Furans / Q000138:chemical synthesis / Q000737:chemistry*; D008968:Molecular Conformation; D009570:Nitriles / Q000138:chemical synthesis / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D010777:Photochemistry
chemical_list: D005456:Fluorescent Dyes; D005663:Furans; D009570:Nitriles; D010455:Peptides