Confocal Laser Scanning Microscopy (CLSM) of decayed wood
IRG/WP 98-10273
Ying Xiao, R N Wakeling, A P Singh
Confocal laser scanning microscopy (CLSM) is currently being used to examine biodegraded wood and is proving to be a useful new technique in this area of work. Non-invasive optical sectioning within a thick specimen (20 - 50 µm), coupled with post image processing techniques allows manipulation of images and 3-D reconstruction from serial sections.
Glutaraldehyde can cause cell autofluorescence, and since wood cell components do not react with it, a relatively simple technique for localising fungal hyphae, using glutaraldehyde as a fixative, was developed. Subsequent use of probes specific for chitin provided superior images of fungal hyphae in wood. High resolution, sequential, 2-D images can be produced to determine the mode of fungal attack within a thick wood specimen in a dynamic way. Voxel 3-D reconstruction of a series of image stacks enabled stereo viewing of objects.