A biomimetic bone-like collagen-hydroxyapatite (Col-HA) composite coating was formed on a surface-treated Ti-6Al-4V alloy substrate via simultaneous collagen self-assembly and hydroxyapatite nucleation. The coating process has been carried out by immersing sand-blasted, acid-etched and UV irradiated Ti-6Al-4V alloy in type I collagen-containing modified simulated body fluid (m-SBF). The surface morphology and phase composition of the coating were characterized using various techniques. More importantly, dual-beam FIB/SEMs with either gallium ion source (GFIB) or xenon plasma ion source (PFIB) were used to investigate the cross-sectional features of the biomimetic Col-HA composite coating in great details. As a result, the cross-sectional images and thin transmission electron microscopy (TEM) specimens were successfully obtained from the composite coating with no obvious damages or milling ion implantations. Both the cross-sectional SEM and TEM results have confirmed that the Col-HA coating demonstrates a similar microstructure to that of pure HA coating with homogeneously distributed elements across the whole cross section. Both coatings consist of a uniform, crack-free gradient structure with a dense layer adjacent to the interface between the Ti-6Al-4V substrate and the coating facilitating a strong bonding, while a porous structure at the coating surface aiding cell attachment. (C) 2018 Elsevier B.V. All rights reserved.