RESUMEN
Supported lipid bilayers (SLBs) have been widely used to provide native environments for membrane protein studies. In this study, we utilized graphene field-effect transistors (GFETs) coated with a fluid SLB to perform label-free detection of membrane-associated ligand-receptor interactions in their native lipid bilayer environment. It is known that the analyte-binding event needs to occur within the Debye length for it to be significantly sensed by an FET sensor. However, the thickness of a lipid bilayer is around 4-5-nm-thick, which is larger than the Debye length of a solution with physiologically relevant ionic strength. There is thus a question of whether an FET sensor can detect the binding event above the bilayer. In this study, we show how the existence of an SLB can influence the effective detection distance and the formation criterion of a fluid and continuous SLB on a graphene surface. We discovered that the water intercalation between the graphene and the underlying silica substrate hinders the SLB formation but is required for the stable electrical recording by a GFET. To verify the existence of a fluid SLB on graphene, which was previously complicated by the graphene fluorescence quenching effect, we developed a modified fluorescence recovery after photobleaching method. In addition, our results showed that SLB coated GFETs can quantitatively detect ligand binding onto the receptors embedded in the SLBs. The comparison of our experimental data with a theoretical model shows that the contribution of the SLB acyl chain hydrophobic region to the screening effect can be negligible and, therefore, that the effective detection region can extend beyond the SLB.
Asunto(s)
Recuperación de Fluorescencia tras Fotoblanqueo/métodos , Grafito/química , Membrana Dobles de Lípidos/química , Transistores Electrónicos , Animales , Biotina/análogos & derivados , Biotina/metabolismo , Bovinos , Colorantes Fluorescentes/química , Ligandos , Fosfatidilcolinas/química , Fosfatidiletanolaminas/metabolismo , Albúmina Sérica Bovina/metabolismo , Estreptavidina/metabolismo , Agua/químicaRESUMEN
The fluorescence enhancement in broadband Cr-doped fibers (CDFs) fabricated by a drawing tower with a redrawn powder-in-tube preform is proposed and demonstrated. The CDFs after heat treatment exhibited Cr4⺠emission enhancement with spectral density of 200 pW/nm, verified by the formation of α-Mg2SiO4 nanocrystalline structures in the core of CDFs. The high fluorescence achievement in the CDFs is essential to develop a broadband CDF amplifier for next-generation optical communication systems.