The specific design and modification of surfaces is of great interest, especially for functional surfaces and medical applications. In order to obtain films on a surface, the layer-by-layer deposition of polyelectrolytes represents a well-established methodology. The alternating deposition of poly(L-histidine) and poly(c-glutamic acid) results in a defined, continuous surface coating that was thoroughly characterized using X-ray photoelectron spectroscopy, Fourier-transform infrared spectroscopy, ellipsometry, X-ray reflectometry, atomic force microscopy, scanning electron microscopy, contact angle, and electrokinetic measurements. Surface charge, film growth, and final thickness were measured and cross-validated. Additionally, the chemical composition and distribution of polyelectrolytes in the layerstack were determined. Finally, the optical parameters were specified and the surface topography was visualized by several methods. These characterizations revealed a coating with embedded spheroids forming from the bottom layers. This rough surface formed by (PLH/PGA)(8) was highly reproducible and might provide unique features for the design of tailored surfaces. (C) 2012 Elsevier Inc. All rights reserved.