In situ analysis of biofilms on historic window glass using confocal laser scanning microscopy

Microbial colonization of the surface of historic glass panels and the subsequent biodeteroration of glass are well documented phenomena. Yet little is known about the composition of this microflora that has to be adapted to low nutrient conditions and a dry environment. The microbial community growing on glass window panels from four different locations and ages ranging from 30 to 600 years was analyzed in situ using confocal laser scanning microscopy with nucleic acid stains and fluorescently labeled rRNA-targeted oligonucleotide probes for the domains Bacteria and Eucarya. A typical biofilm of the studied glasses displayed a total thickness of approximately 10-60 μm. Microbial colonization of the glass surface was heterogeneous at 0.8-7% areal coverage. The dominant microbial group belonged to the filamentous fungi. A different attached microflora was found only on one glass surface. This sample was sparsely colonized with areal coverage of 0.8% and a thickness of 10-20 μm; the biofilm consisted of single bacterial cells and microcolonies. Chemical composition and durability of the glass samples and availability of an additional organic layer were important factors influencing the extent of microbial growth. Information about the thickness and microbial composition of biofilms offer an essential background to optimize cleaning procedures or conservation strategies for stained glass windows.