Poly(hexafluoropropylene-co-tetrafluoroethylene) (FEP) has been widely used in biotechnology because of its unique surface properties and biocompatibility. Recent work from our group has shown that plasma discharge-modified FEP can be used as the substratum for development of a very stable immunosensor. This result has prompted us to study further this new surface under ambient conditions. In this paper, we report on the covalent immobilization of a pyrene residue (-Py) onto FEP-APS (FEP-aminopropyl silane) surfaces and the characterization of FEP-APS-Py using steady-state and time-resolved fluorescence spectroscopy. Among the immobilization schemes tested, we found that the covalent coupling of pyrene-sulfonyl chloride to FEP-APS is the easiest and yields the most photostable FEP-APS-Py derivative. Steady-state emission spectra of FEP-APS-Py in contact with H2O, β-cyclodextrin (β-CD), and sodium dodecylsulfate (SDS) aqueous solutions differ considerably from those of Py-SO3 in solution. Time-resolved fluorescence spectroscopy of FEP-APS-Py demonstrates that the decay kinetics are strongly affected by the presence of ionic quenchers and molecular oxygen, as well as β-CD and SDS. The results are consistent with the suggestion that the APS-Py moiety undergoes a slow time-dependent reconfiguration at the FEP/APS interface.
Time-resolved fluorescence spectroscopy. Applications to calmodulin