Polarization optical analysis of the surface structures of various fungi.

Vicinal hydroxyl groups of the sugar compounds sialic acid and 9-O-acyl sialic acid can be visualised for polarization optical analysis on the surface of different fungi using several topo-optical reactions. We investigated the presence of these molecules in cultures of Cryptococcus neoformans (heterogeneous form), Saccharomyces cerevisiae, Candida albicans, Candida glabrata, Candida krusei and Candida tropicalis by topo-optical reactions. Additionally, we examined brain and stomach tissues of patients with infections by C. neoformans and C. albicans, respectively. The results suggest a highly fashioned orientation of the sugar chains on the fungal surface. Terminal sialic- and O-acyl sialic acid residues are permanently present and orientated in a highly specific way in the cell wall of fungi. Based on the polarization optical analysis after the ABT-r (anisotropic PAS-r), the linear oriented hydroxyl groups of the sugar molecules are localized either perpendicular or parallel to the surface coat, depending on the species. According to the orientation of the vicinal hydroxyl groups, the oligosaccharide chains are orientated vertically. The capsule of the heterogeneous form of C. neoformans presented an especially strong metachromatic reaction and anisotropy. It is especially remarkable that the sterical orientation of sugar chains, and the terminal sialic acid and 9-O-acyl sialic acid molecules, was opposite in the inner and outer layer of the capsule.

Topo-optical investigations on the surface of bacterial cells during the phagocytosis of Klebsiella pneumoniae in mouse.

Polarisation optical methods provide the means to perform sub-microscopic investigations on structures containing spatially highly ordered molecules, for example the cell envelope of prokaryotic cells. Such structures can evoke birefringence, which can be enhanced or modified by different dyes or reagents, thus providing the possibility of a more specific investigation of the composition and structure of bacterial surface compounds. Klebsiella pneumoniae synthesises sterically different carbohydrate-rich structures, including those of the outermost capsular polysaccharide, the polysaccharide somatic antigen of the lipopolysaccharide molecule and the peptidoglycan layer of the cell wall. In the study reported here, the nature and intensity of topo-optical activity of these structures was analysed using the aldehyde-bisulphite-toluidine blue reaction, sialic acid topo-optical reactions and chlorpromazine-eosin charge transfer reactions. Furthermore, a mouse intraperitoneal model was used to analyse alterations in topo-optical characteristics of bacteria during phagocytosis. Both encapsulated and non-encapsulated bacterial cells changed their original pattern and orientation of birefringence after being phagocytosed.