Title
Amine-Reactive Fluorene Probes: Synthesis, Optical Characterization, Bioconjugation, And Two-Photon Fluorescence Imaging
Abstract
With the increasing demand for confocal and two-photon fluorescence imaging, the availability of reactive probes that possess high two-photon absorptivity, high fluorescence quantum yield, and high photostability is of paramount importance. To address the demand for better-performing probes, we prepared two-photon absorbing amine-reactive fluorenyl-based probes 2-(9,9-bis(2-(2-methoxyethoxy)ethyl)-2-isothiocyanato-9H-fluoren-7-yl) benzothiazole (1) and 2-(4-(2-(9,9-bis(2-(2-ethoxyethoxy)ethyl)-2- isothiocyanato-9H-fluoren-7-yl)vinyl)phenyl)benzothiazole (2), incorporating the isothiocyanate as a reactive linker. Probe design was augmented by integrating high optical nonlinearities, increased hydrophilicity, and coupling with reactive functional groups for specific targeting of biomolecules, assuring a better impact on two-photon fluorescence microscopy (2PFM) imaging. The isothiocyanate (NCS) derivatives were conjugated with cyclic peptide RGDfK and Reelin protein. The study of the chemical and photophysical properties of the new labeling reagents, as well as the conjugates, is described. The conjugates displayed high chemical stability and photostability. The NCS derivatives had low fluorescence quantum yields, while their bioconjugates exhibited high fluorescence quantum yields, essentially "lighting up" after conjugation. Conventional and 2PFM imaging and fluorescence lifetime imaging (FLIM) of HeLa, NT2, and H1299 cells, incubated with two-photon absorbing amine-reactive probe (1), RGDfK-dye conjugate (7), and Reelin-dye conjugate (6), was demonstrated. © 2008 American Chemical Society.
Publication Date
12-17-2008
Publication Title
Bioconjugate Chemistry
Volume
19
Issue
12
Number of Pages
2559-2567
Document Type
Article
Personal Identifier
scopus
DOI Link
https://doi.org/10.1021/bc800415t
Copyright Status
Unknown
Socpus ID
58149083267 (Scopus)
Source API URL
https://api.elsevier.com/content/abstract/scopus_id/58149083267
STARS Citation
Morales, Alma R.; Schafer-Hales, Katherine J.; Marcus, Adam I.; and Belfield, Kevin D., "Amine-Reactive Fluorene Probes: Synthesis, Optical Characterization, Bioconjugation, And Two-Photon Fluorescence Imaging" (2008). Scopus Export 2000s. 9410.
https://stars.library.ucf.edu/scopus2000/9410