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The page below is a sample from the LabCE course Histology Special Stains: Connective Tissue. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Gordon and Sweet's Silver Staining - 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% iron alum solution
10 minutes
Running water wash1 minute

Distilled water3 changes
Working ammoniacal silver solution
7 dipsShake off excess solution
Distilled water
2 changes, 3 quick dips each
10% formalin30 secondsTissue will turn gray black
Distilled water3 changes
0.5% gold chloride
1 minuteOptional step; tones/lightens the gray black silver to light gray
Distilled water
3 changes
5% hypo1 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 with 95% and absolute alcohols. Follow with two changes of xylene and then coverslip.
    Expected results:
    • Reticular fibers - Black (with linear pattern)
    • Nuclei - Red
    • Other tissue elements - Pinkish-red