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Old 02-06-04, 11:55 AM   #49 (permalink)
Scales Zoo
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Join Date: Mar-2007
Posts: 1,867
Thanks Dark Angel.

And from another link already posted in this thread

To make a more rapid diagnosis, cytologic smears can be made from biopsies and stained using a modified hematoxylin and eosin (H&E) technique:

1. Fix 1 minute in 10% neutral buffered formalin
2. Stain 3 minutes in Harris hematoxylin
3. One dip into acid alcohol
4. Briefly wash in running water
5. Dip into 0.5%ammonia to blue the nuclei
6. Wash in running water
7. Counterstain in eosin for 40 seconds
8. Dehydrate in a series of alcohols: 95%-100%-100%
9. Place in xylene and mount as with a paraffin embedded section

I have found this staining far superior than using Wright-Giemsa staining. With Wright Giemsa staining the inclusions stain basophilic and may not be readily recognizable. With H&E staining, inclusions stain exactly the same as in tissues embedded in paraffin, sectioned and stained with H&E. It must be remembered that absence of inclusions in the biopsies does not necessarily mean the snake is free of IBD. In some cases inclusions may only be seen in the central nervous system, and in those cases may be few in number. The diagnosis is only as good as the portion sampled and biopsies routinely represent a relatively small portion of the entire tissue.

If inclusions are identified in any cells, euthanasia is recommended.
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