A new approach to study local corrosion processes on steel surfaces by combining different microscopic techniques

Corrosion studies of materials on the micro or even nano-scale level are cumbersome due to instrumental limitations and handling procedures. If biological processes are involved the spatial resolution is even more important and sample preparation is usually the limitation. Attachment of bacteria on stainless steel surface is a complex interfacial process including interactions of bacterial cells and bacterial extracellular polymeric substances with the surface. To overcome the limitations in sample preparations and resolution we present a new stainless steel sample holder to switch among epifluorescent microscope (EFM), AFM and SEM at exactly the same position. Exemplary bacterial accumulation was studied by staining the bacterial DNA with a fluorescent dye over time. It was possible to distinguish among bacteria and other surface characteristic such as deformations or grain structures. Also surface topographic features such as roughness at the grain boundaries and deposits were quantified. All three techniques complement one another in the way that AFM is a high-resolution technique that does not allow to distinguish directly bacterial cell structures, whereas EFM offers excellent bacterial identification based on staining at a low resolution that can complement AFM images. Application of SEM in the last step will reveal inclusions and grain structure and combined with EDX gives the composition of the substrate, inclusions and corrosion deposit. The combination of the three high-resolution techniques enables a more detailed understanding of surface phenomena. The method itself is quite elegant and easy to handle which is an important aspect in materials research, especially when a high sample throughput is needed. © 2012 Elsevier B.V. All rights reserved.