Nonlinear optical studies of self-assembled monolayers (SAM) silica-SAM-water interface probed with second harmonic generation

Second harmonic generation (SHG) is a successful and widely used technique for the study of surfaces and surface phenomena. We present a novel technique using second harmonic generation from oriented water molecules in the Gouy-Chapman diffuse layer at the alkylsiloxane and biomolecular self assembled monolayer (SAM) interface with water to measure distance between the solid surface and the average location of the oriented water in the diffuse layer. This distance is manifest in the SHG angular dependence profile from the relative phases of the second harmonic light generated at the diffuse layer and at the solid surface. Distances of one nanometer can be distinguished. Values for the diffuse layer potential, diffuse layer decay length, magnitude and phase of the nonlinear susceptibility were obtained. This in situ probe is universally applicable for organic adsorbates which in general will push the diffuse layer away from the solid surface. The organic layer thickness can be used to obtain the adsorption fraction. From this and an understanding of the likely chemistry, the orientation of the molecules can be inferred. We have demonstrated this technique on three molecular systems: hydrophobic self assembled monolayers of methoxysilane molecules of varying hydrocarbon chain length, self assembled monolayers of streptavidin glycoproteins and the combined streptavidin-biotinylated antibody monolayer. In the methoxysilane monolayers a relationship between hydrophobicity and molecular orientation was observed. The thickness of the streptavidin monolayer was determined to be 5.6 nm. This is strikingly close to the length of the of the streptavidin molecule which implies a close packed monolayer of streptavidin molecules. The average height of the antibodies was determined to be 10.9 nm or about two thirds the height of an antibody molecule. This too confirms a monolayer and allows for good approximation of surface coverage. This method does nothing to disturb or alter the structure or function of the monolayers during measurement. No physical probe or chemical reagent is ever introduced into the monolayer or its water environment. This is of immense value to the study of multi-layer systems like immobilized antibodies. The surface can be accurately characterized at every phase of monolayer development without compromising the integrity of the developing system or the measurements themselves.