Gordon and Sweet's Silver Stain - Staining Protocol

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Gordon and Sweet's Silver Stain - Staining Protocol

Sample type required: Deparaffinized and rehydrated tissue section on positively (+) charged slides

Preferred fixative: 10% neutral buffered formalin (NBF)

Control: Normal liver

ReagentTimeTechnical Notes
1% potassium permanganate solution5 minutes
Running water wash 2 minutes

1% oxalic acid wash Until tissue is colorless

Distilled water3 changes
2% ferric ammonium sulfate solution
at least 15 minutes
Running water wash1 minute

Distilled water3 changes
Working ammoniacal silver solution
5 minutesShake off excess solution
Distilled water
2 changes, 3 quick dips eachRinse well
10% formalin2 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 water3 minutes
0.5% gold chloride 30 seconds to 1 minute
Distilled water
3 changes
5% sodium thiosulfate1 minute
Running water wash1 minute

Nuclear fast red solution
5 minutes
Running water wash
1 minute
Red color will remain in tissue

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