title: Two-photon fluorescence vascular bioimaging with new bioconjugate probes selective toward the vascular endothelial growth factor receptor 2.
abstract: We report the synthesis and characterization of two amine reactive fluorescent dyes with efficient two-photon absorption (2PA) properties and high fluorescence quantum yields. Bioconjugation of these dyes with the DC-101 antibody proved to be useful for selectively imaging the vascular endothelial growth factor receptor 2 (VEGFR-2) in cells expressing this receptor in vitro and in "whole" mounted excised tumors (ex vivo) by two-photon fluorescence microscopy (2PFM). The penetration depths reached within the tumors by 2PFM was over 800 μm. In addition, the concentration of dye required for incubation of these bioconjugates was in the picomolar domain, the probes possessed very good photostability, and the 2PFM setup did not require any additional means of increasing the collection efficiencies of fluorescent photons to achieve the relatively deep tissue imaging that was realized, due, in large part, to the favorable photophysical properties of the new probes.
authors.affiliation: Department of Chemistry, The College of Optics and Photonics, University of Central Florida, Orlando, Florida 32816, United States.
authors.forename: Carolina D
authors.initials: CD
authors.lastname: Andrade
authors.forename: Ciceron O
authors.initials: CO
authors.lastname: Yanez
authors.forename: Hyo-Yang
authors.initials: HY
authors.lastname: Ahn
authors.forename: Takeo
authors.initials: T
authors.lastname: Urakami
authors.forename: Mykhailo V
authors.initials: MV
authors.lastname: Bondar
authors.forename: Masanobu
authors.initials: M
authors.lastname: Komatsu
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Belfield
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D018827:Carcinoma, Lewis Lung / Q000175:diagnosis*; D002460:Cell Line; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D057054:Molecular Imaging; D013552:Swine; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D018796:Immunoconjugates; D040301:Vascular Endothelial Growth Factor Receptor-2