Visualisation of Native Surfaces by Two-Step Molding

Abstract One of the most important tasks of microscopy is to provide information about structures in their natural state. Since most life science microscopy procedures require fixation, dehydration, drying, and sectioning, diverse artefacts are unavoidable. It is also possible to dehydrate the water content of a specimen by freeze-drying. However, even relatively stable surface structures are changed during such treatments. Methods of liquid substitution and freezing-substitution show good results for biological specimens with a waxy solid coverage (Ensikat and Barthlott, 1993) and soft specimens in a mechanically deformed state (Gorb et al., 2000). However, all these methods have certain restrictions for some types of specimens. For example, there are numerous biological surfaces covered with secretory fluids. Some specimens bear solid waxy coverings, which can be partly dissolved and washed out in organic solvents, such as ethanol, acetone, or propylenoxide. Often one would like to visualise dynamic processes (growth, deformation, or crystallisation) under environmental conditions, but at the high resolution in the SEM.