Protein–membrane interactions investigated with surface-induced fluorescence attenuation

Research on protein-membrane interactions has been undeveloped due to the lack of proper techniques to detect the position of proteins at membranes because membranes are usually only about 4-nm thick. We have recently developed a new method named surface-induced fluorescence attenuation （SIFA） to track both vertical and lateral kinetics of a single labelling dye in supported lipid bilayers. It takes advantage of strong interaction between a light-emitting dye and a partially reflecting surface. By applying the technique to membrane proteins being fluorescently labelled at different residues, here we show that SIFA can measure not only the insertion depth of a dye inside a lipid bilayer, but also the position of a dye in solution near the surface. SIFA can therefore be used to study membrane proteins of various types.

Segmental dynamics near the chain end of polystyrene in its ultrathin films:a study by single-molecule fluorescence de-focus microscopy

Rotational motion of fluorophores chemically attached to polystyrene chain-ends in ultra-thin films on solid substrates was studied by single-molecule fluorescence de-focus microscopy.The collective feature of the rotational motion was found and evidenced by the sharp change of the population of fluorophores undergoing rotational motion within a very narrow temperature range(named as the changing temperature,T c).The T c value was found to depend on film thickness and interfacial chemistry and the variation of the T c value is also dependent on the molecular weight of the polymer.The results demonstrate that the spatial confinement effect enhances the segmental mobility near the polymer chain-ends while the interfacial attraction restricts the segmental motion inside the thin film.

Near-Field Spot for Localized Light-Excitation of a Single Fluorescent Molecule

Zero-mode waveguides have become important tools for the detection of single molecules.There are still,however,serious challenges because large molecules need to be packed into nano-holes.To circumvent this problem,we investigate and numerically simulate a novel planar sub-wavelength 3-dimension(3D)structure,which is named as near-field spot.It enables the detection of a single molecule in highly concentrated solutions.The near-field spot can produce evanescent waves at the dielectric/water interface,which exponentially decay as they travel away from the dielectric/water interface.These evanescent waves are keys for the detection of fluorescently tagged single molecules.A numerical simulation of the proposed device shows that the performance is comparable with a zero-mode waveguide.Additional degrees-of-freedom,however,can potentially supersede its performance.

When does a diblock copolymer probe the interfacial rheological effect?

Lateral diffusion of diblock copolymer residing on the interfaces between two immiscible liquids is investigated at single molecular level. Fluorescence correlation spectroscopy was used to study the diffusion of fluorescence-labeled diblock copolymer,polystyrene-b-polyisoprene, at the interface formed between two immiscible liquids. The interfaces are formed between N,N-dimethylformamide(DMF) and a few immiscible liquids, n-alkane and polyisoprene. Interfacial diffusion coefficient of the diblock copolymer probe is found to decrease monotonously with the increase of the molecular length of the interface constituting liquids. The decrease of diffusion coefficient follows the prediction by Einstein relation using the viscosity of the constituting liquids as the variables only for interfaces between DMF and very small n-alkanes. For interfaces formed between DMF and bigger alkanes and especially between DMF and polyisoprene, the diffusion coefficient is much higher than the calculated value,indicating that the probe molecule starts to probe the much less viscous interfacial region because the interfacial width gets larger,whose thickness is comparable to the molecule size of the liquids.