Sequential SEM imaging of microbial calcite precipitation consolidation treatment

Cultural heritage built from limestone is prone to deterioration by chemical weathering, a natural process, that is enhanced by pollution. There are many historic monuments built from calcareous rocks that suffer from deterioration, and thus there have been a number of approaches over the last few decades to consolidate these types of rocks and surfaces. Using natural biological processes by fostering the activity of calcite-producing bacteria, also referred to as biomineralization, is one strategy that has also been commercialized. The base of proving the effectiveness of any surface treatment is the observation of the surface at sequential stages before and after treatment, as well as after exposure to weathering. Due to the heterogeneity of natural materials and processes, our aim was to observe identical test areas at the micron scale throughout the observation period. In order to achieve this on a tungsten SEM, we employed a beam deceleration accessory that allowed low kV imaging on non-conductive surfaces at a sufficiently high image resolution with a modified sample holder accommodating drill cores of 25 mm diameter and up to 15 mm height. The presented method is capable of producing time-sequenced images on the same test area on natural rock surface samples without manipulation for imaging purposes. This offers interesting perspectives for effective documentation of such processes in various fields.