Sample type required: Deparaffinized and rehydrated tissue section on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Normal liver
| Reagent | Time | Technical Notes |
| 1% potassium permanganate solution | 5 minutes |
|
| Running water wash | 2 minutes |
|
| 1% oxalic acid wash | Until tissue is colorless |
|
| Distilled water | 3 changes |
|
| 2% ferric ammonium sulfate solution | At least 15 minutes |
|
| Running water wash | 1 minute |
|
| Distilled water | 3 changes |
|
| Working ammoniacal silver solution | 5 minutes | Shake off excess solution |
| Distilled water | 2 changes, 3 quick dips each | Rinse well |
| 10% formalin | 2 minutes | The tissue will turn dark brown to black. Note: If sections do not turn dark brown/black immediately, rinse slides and repeat the impregnation step, checking the working silver solution for reactivity and proper composition. |
| Distilled water | 3 minutes |
|
| 0.5% gold chloride | 30 seconds to 1 minute |
|
| Distilled water | 3 changes |
|
| 5% sodium thiosulfate | 1 minute |
|
| Running water wash | 1 minute |
|
| Nuclear fast red solution | 5 minutes |
|
| Running water wash | 1 minute | The red color will remain in the tissue. |
Post staining procedure:
The tissue section should be rinsed well in distilled water and dehydrated through graded alcohols (beginning with 95%). Two changes of xylene should follow, followed by a coverslip.
Expected results:
- Reticular fibers - Black (with linear pattern)
- Nuclei - Red
- Other tissue elements - Pinkish-red
