Amine-reactive fluorine-based probes: Synthesis, optical characterization, bioconjugation, and two-photon fluorescence imaging

ORGN 434

Alma R. Morales, amorales@mail.ucf.edu1, Kevin D. Belfield, belfield@mail.ucf.edu2, and Zhen-Li Huang, huangz@creol.ucf.edu2. (1) Department of Chemistry, University of Central Florida, 4000 Central Florida Boulevard, P.O. Box 162366, Orlando, FL 32816-2366, (2) Department of Chemistry and CREOL, College of Optics and Photonics, University of Central Florida, 4000 Central Florida Blvd., P.O. Box 162366, Orlando, FL 32816-2366
We have been focused on developing select fluorene derivatives exhibiting desirable properties: high fluorescence, photostability, two-photon absorption cross-section, and low cytotoxicity to enhance their functionality and/or specificity for biological targets and to increase the solubility of these compounds in aqueous environments. Our efforts, directed toward preparing reactive fluorescent reagents, involve the synthesis of amine-reactive fluorene derivatives. Our strategy consists of building fluorene-based structure containing the isothiocyanate functionality, capable of forming a stable linkage with biomolecules containing amino groups. The synthetic details of amine reactive probes, a model adduct, along with a model bioconjugate, and its single-photon spectral characteristics, as well as the two-photon absorption cross sections will be discussed. Additionally, we demonstrate that the fluorenyl isothiocyanates react spontaneously with a protein in HeLa cells, forming a highly fluorescent bioconjugate, readily detected by single-photon and two-photon fluorescence microscopic imaging.
 

New Reactions and Methodology, Total Synthesis, Materials, Devices and Switches, Lipids, Nucleotides and Mimetics
8:00 PM-10:00 PM, Tuesday, March 27, 2007 Hyatt Regency Chicago -- Riverside Center, Poster

Division of Organic Chemistry

The 233rd ACS National Meeting, Chicago, IL, March 25-29, 2007