PAS Diastase Stain Protocol

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

 

An Expert Histology Service


Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s