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