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Summary This work reports an improved preparation procedure for acquiring SEM images for three-dimensional (3D) reconstruction. The images are acquired consecutively after serial grinding and polishing. Ugelstad beads are used as landmarks for registration purposes and for determining the thickness of the abraded sections. An estimation of the bead sizes necessary for suitable thickness quantification at the corresponding spatial resolution and uncertainty is given. Shape-based interpolation is used for filling the gaps between the cross-sections. An indication of the distance between cross-sections necessary for good interpolation is also given. An example of a 3D reconstructed paper volume is presented. The method is suitable for preparation of fibre and paper as well as other materials.

Cyanoacrylic glue (Eastman 910) was used to affix small pieces of nasal scrapings to lens paper immediately before fixation in the glutaraldehyde. The lens paper not only served to hold specimens together so that they were not lost during tissue processing, but also functioned as a 'landmark' for the specimens, so that specimens could be oriented in a specific manner during embedding and subsequent sectioning.