Sample type required: Deparaffinized and rehydrated tissue section (3–5 microns) on positively (+) charged slides
Preferred fixative: Zenkers; however, 10% neutral buffered formalin (NBF) should be used due to removal of mercury from labs
Control: Skeletal or cardiac muscle
| Reagent | Time | Technical Notes |
| Zinc chloride | 5 minutes
(in heated solution) | They are used as a secondary fixative and mordant. Microwave the solution for 20 seconds on high power before placing the slides inside. Microwave solutions should not be reused. |
| Running water wash | 5 minutes | Ensure that ALL traces of the reagent have been removed. |
| 0.25% potassium permanganate | 5 minutes |
|
| Running water wash | 5 minutes |
|
| 5% oxalic acid | 1 minute | Tissue should be bleached/colorless after exposure |
| Running water wash | 10 minutes |
|
| Distilled water | 3 changes | Discard the solution after use. |
| PTAH staining solution | Overnight at room temperature |
|
Post staining procedure:
Dehydrate quickly through two changes of 95% and 100% alcohol, followed by two changes of xylene, then cover-slip.
Expected results:
- Cross-striations - Blue
- Nuclei - Blue
- Collagen - Red-brown
- Elastic fibers - Purplish
