title: Microwave-assisted synthesis of thiophene fluorophores, labeling and multilabeling of monoclonal antibodies, and long lasting staining of fixed cells.
abstract: We report the expedient microwave-assisted synthesis of thiophene based 4-sulfo-2,3,5,6,-tetrafluorophenyl esters whose molecular structure was engineered to achieve blue to red bright fluorescence. The reactivity toward monoclonal antibodies of the newly synthesized fluorophores was analyzed in comparison with that of the corresponding N-succinimidyl esters. Single-fluorophore and multiple-fluorophore labeled antibodies were easily prepared with both types of esters. Multiple-fluorophore labeling with blue and orange emitting fluorophores resulted in white fluorescent antibodies. Thiophene based fluorophores displayed unprecedented fluorescence stability in immunostaining experiments. First-principles TD-DFT theoretical calculations helped us to interpret the behavior of fluorescence emission in different environments.
authors.affiliation: Consiglio Nazionale Ricerche CNR-ISOF, Via Gobetti 101, 40129 Bologna, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Zambianchi
authors.forename: Francesca
authors.initials: F
authors.lastname: Di Maria
authors.forename: Antonella
authors.initials: A
authors.lastname: Cazzato
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gigli
authors.forename: Manuel
authors.initials: M
authors.lastname: Piacenza
authors.forename: Fabio
authors.initials: F
authors.lastname: Della Sala
authors.forename: Giovanna
authors.initials: G
authors.lastname: Barbarella
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D002460:Cell Line; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D006801:Humans; D008214:Lymphocytes; D008872:Microwaves; D013194:Staining and Labeling / Q000379:methods*; D013876:Thiophenes / Q000737:chemistry*; D016707:Tissue Fixation
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D013876:Thiophenes