Structural studies of membranes and surface layers up to 1,000 A thick using X-ray standing waves.

The X-ray standing wave (XSW) method, developed in the 1960s, was used originally to determine heavy atom positions in and on silicon and germanium single crystals. An X-ray standing wave generated by the interference of coherent incident and reflected beams excites X-ray fluorescence from the heavy atom, the intensity of which as a function of incident angle provides an indication of the atom's distance from the X-ray reflecting surface. The availability of X-ray mirrors and the ability to prepare layered synthetic microstructures has made possible the study of biologically relevant structures using the XSW technique on length scales of typically tens to hundreds of ångströms, allowing heavy atoms in such structures to be located with ångström or subångström resolution. Many model biological systems (such as Langmuir-Blodgett films, which mimic membranes) require access to still larger scales, but it is not obvious that an XSW will remain coherent over such length scales. Here we report studies of a lipid multilayer system using the XSW method, in which we have been able to locate the metal atoms in a zinc arachidate bilayer with ångström resolution at a distance of almost 1,000 A above the surface of a gold mirror. Our results indicate that the XSW technique should be useful for structural studies of supramolecular aggregates, receptor-ligand interactions and multi-membrane stacks, in which length scales of this order are encountered.

Diffuse-double layer at a membrane-aqueous interface measured with x-ray standing waves.

The ion distribution in an electrolyte solution in contact with a charged polymerized phospholipid membrane was directly measured with long-period x-ray standing waves. The 27-angstrom-thick lipid monolayer was supported on a tungsten/silicon mirror. X-ray standing waves were generated above the mirror surface by total external reflection of a 9.8-kiloelectron volt x-ray beam from a synchrotron undulator. The membrane surface, which contained negatively charged phosphate headgroups, was bathed in a dilute ZnCl2 solution. The concentration of Zn2+ in the condensed layer at the membrane surface and the Zn2+ distribution in the diffuse layer were measured as a function of headgroup charge. The Debye length of the diffuse layer varied between 3 and 58 angstroms. The results qualitatively agree with the Gouy-Chapman-Stern model.