For histological examination, tissue samples will be collected and fixed instantly in 10% buffered formalin for up to 48hs.

Therefter, the samples will be dehydrated sequentially in 70% alcohol, 80% alcohol, 95% alcohol and then finally in absolute alcohol. Clearing of the tissue samples will then be performed by placing them in xylene in order to remove retained alcohol within the tissue spaces. Therafter, the tissue samples will be fixed into liquid paraffin wax which will be placed into a vacuum oven connected to a suction pump. This will remove air within the tissue while the liquid paraffin goes into occupy the vacuum created thereby impregnating the tissue. The impregnated tissue will then placed into an embedding iron which will be filled with liquid paraffin. An embedding ring will then be placed into the embedding iron already containing the tissue sample. The paraffin will be allowed to solidify into a block. The embedded tissue block will be sectioned at 5 10 m thickness using a microtome knife and microtome machine in order to produce the tissue section. A drop of egg albumin-glycerine mixture will be used to smear the surface of the slide. The tissue section will then be placed on the smeared surface. The slide will then be flooded with 20% alcohol. The slide will be placed on a hot plate until the tissue sections is flattened and glued to the surface of the slide. This process ensures that the tissue section is not wrinkled or detached from the slide during the process of staining. The 20% alcohol will be then drained from the slide and the slide will be dried by placing it into a hot oven. Haematoxylin-Eosin staining will then be undertaken as recommended by..........; the dried slide will be placed into xylene for 5mins to clear the paraffin from the tissue section and then dipped sequentially through absolute alcohol and then 95% alcohol, 80% alcohol and
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finally 70% alcohol. The slide will then be rinsed with tap water for 5mins. Thereafter, the slide will be stained with Haematoxylin for 15 mins and then rinsed very well in tap water. The slide will then be decolourised with 1% acid alcohol and then rinsed with tap water for 20 mins to stop the action of the acid alcohol. Thereafter,the slide will be dipped into Eosin for 1 2mins after which it will be rinsed very well in tap water. The slide will then be dipped sequentially through 70% alcohol, 80% alcohol, 95% alcohol and absolute alcohol and then into xylene. The slide will be removed from xylene and a drop of DPX mountant will be placed on the tissue section of the slide. A coverslip will then be placed on the drop of mountant after which the slide will be taken to the oven for drying. After drying the slide, it will be examined using a light microscope at magnifications of x10, x40 and with oil immersion at x100. Significant observations will be recorded.

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