Sample Type Required: Deparaffinized and re-hydrated tissue sections on positively charged slides.
Fixative:10% Neutral Buffered Formalin
| Step | Reagent | Time | Technical Notes |
| 1 | 12% Acetic Acid Solution | Brief Rinse |
|
| 2 | Working Colloidal Iron Solution (10mL Stock Colloidal Iron, 18mL Distilled Water and 12mL Glacial Acetic Acid) | 1 Hour |
|
| 3 | 12% Acetic Acid Solution | 3 Changes x 3 Minutes Each |
|
| 4 | Ferrocyanide-Hydrochloric Acid Solution (25mL Potassium Ferrocyanide, 2% and 25mL Hydrochloric Acid, 2%) | 20 Minutes |
|
| 5 | Running Tap Water | 5 Minutes |
|
| 6 | Nuclear Fast Red Solution | 5 Minutes |
|
| 7 | Running Tap Water | 1 Minute | Clouding will result when slides are placed in the alcohols if slides are not washed well after Nuclear Fast Red staining. |
Expected Results
- Acid Mucins and Sialomucins = Deep Blue
- Nuclei = Pink to Red
- Cytoplasm = Pink
Post Staining Procedure: Tissue sections should be rinsed well in distilled water, dehydrated with 95% and absolute alcohols, cleared and cover-slipped.
