If you are going to do a PAS stain then you might as well run a PAS with diastase also.

Safety equipment: Work under a hood with lab coat, gloves, and glasses.

1. Two sets of slides and 2 control slides are used
2. De-paraffin slides in xylene (1) —————— 2 minutes (re-use)
3. De-paraffin slides in xylene (2)—————— 2 minutes (re-use)
4. Clear slides in 100% alcohol——————— 2 minutes
5. Clear slides in 100% alcohol——————— 2 minutes
6. Hydrate slides in 95% alcohol——————- 2 minutes
7. Hydrate slides in running water—————– 5 minutes
8. Set 1 (with a control) in 80% alcohol ———— 60 minutes
9. Set 2 (with a control) in diastase of malt——— 60 minutes
10. Combine both sets of slides for remaining steps
11. Wash in running water ————————– 10 minutes
12. 0.5% periodic acid ——————————- 5 minutes (re-use)
13. Schiff’s reagent at 37C————————— 15 minutes (re-use)
14. Wash in running water ————————– 10 minutes
15. If staining for glycogen, Hematoxylin ———— 20 seconds (re-use)
16. Dehydrate in 95% alcohol———————— 30 seconds
17. Dehydrate in 100% alcohol———————- 1 minute
18. Dehydrate in 100% alcohol———————- 2 minutes
19. Dehydrate in 100% alcohol———————- 2 minutes
20. Clear in xylene (3)——————————- 2 minutes (re-use)
21. Clear in xylene (4)——————————- 5 minutes (re-use)

Results:         Glycogen, mucin, fibrin, or thrombi, colloid droplets, hyaline of arteriosclerosis, hyaline deposits in glomeruli, granular cells in the renal arterioles where preserved, most basement membranes, colloid of pituitary stalks and thyroid, amyloid infiltration may show a positive reaction – Rose to Purplish Red

                        Nuclei – Blue

 

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