When used in conjunction with digital image processing techniques, confocal laser scanning microscopy (CLSM) enables non-invasive optical sectioning, allowing micromorphologies of wood decay to be examined at any depth within a relatively thick (0.05-0.1 mm) wood specimen without incision. In this study, the use of specially tailored multi-fluorescent staining techniques with CLSM produced new information concerning spatial relationships between fungi and bacteria and the wood substrate, particularly in regard to their 3D characteristics. Glutaraldehyde fixation and a chitin fluorescent probe were used to locate fungal hyphae in wood. Bacteria colonising wood were examined using a fluorescent phospholipid probe. By counterstaining wood with this probe and a fluorescent dye specific for Gram-positive bacteria, it was possible to clearly distinguish Gram types through simultaneous, multichannel fluorescent CLSM imaging. The combination of glutaraldehyde fixation and phospholipid probing proved to be reliable for detecting wood-degrading bacteria in wood cell walls.
Light and electron microscopic observations were made of Pinus radiata wood treated to a high retention of copper-chrome-arsenate (CCA), which had failed after approximately 11 years in Service. The predominant wood cell wall degradation patterns seen under the light microscope and scanning electron microscope äs channels and shallow depressions and which were probably responsible for failure of the pole were shown by transmission electron microscopy to be caused by erosion bacteria. Although this type of wood decay is known, the predominance of attack of deeper regions of the pole by erosion bacteria was unusual and significant in terms of the ecology of wood degradirig bacteria. Soft-rot was seen in outer regions of the wood at the groundline but was very rareiy seen in deeper regions.
MicroscopyThe pole had dried out on arrival at the laboratory. Small Chips of wood, 5 mm thick in a radial direction were removed with a chisel at the surface, and at depths of 20, 40, 60 and 80mm, across the diameter of discs. Transverse, radial and.tangential sections were Holzforschung / Vol. 48 / 1994 / No. 6
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