Gordon and Sweet's Stain for Reticulin
Classification: connective tissue stain
Mechanism of staining: argyrophil metallic impregnation
Purpose: stain reticulin
Control tissue: liver, spleen, lymph nodes
Properly Stained Slide
Red: nuclei
Lt. Red: cytoplasm
Black: reticulin
REAGENT | PURPOSE | MECHANISM OF STAINING | SOURCE OF ERROR |
Acidified potassium permanganate | Oxidation | Exposes reactive aldehyde groups | Omitted: No aldehyde groups will be produced and no reticulin fibers will be demonstrated. |
Too short: No aldehyde groups will be produced and no reticulin fibers will be demonstrated. | |||
Too long: Over oxidation of aldehyde groups and no demonstration of reticulin fibers. Background staining. | |||
Oxalic acid | Bleaching | Removes brown discoloration | Omitted: Discoloration in counterstain |
Too short: No effect. | |||
Too long: No effect. | |||
Iron alum | Sensitization | Forms complexes at the aldehyde groups | Omitted: Staining of reticulin with silver stain would not occur. |
Too short: Weak staining of reticulin. | |||
Too long: No effect. | |||
Ammoniacal silver | Impregnation | Silver salt and ferric complex exchange ions | Omitted: No reticulin demonstrated. |
Too short: Underimpregnation. | |||
Too long: Overimpregnation. | |||
10% Formaldehyde | Reduction | Produces a visible black metallic precipitate | Omitted: No reticulin demonstrated. |
Too short: Weakly stained reticulin. | |||
Too long: No effect. | |||
Gold chloride | Tones collagen | Turns collagen from golden to lavender/grey. | Omitted: Collagen will remain golden- brown. Toning is optional. |
Too short: Collagen will remain golden-brown. | |||
Too long: Reticulin will stain weakly when gold replaces silver. | |||
Sodium thiosulphate (hypo) | Fixation | Ion exchange of silver with gold | Omitted: Non-specific blackening of slide over time due to remaining unreduced silver. |
Too short: Some non-specific blackening of slide over time. | |||
Too long: No effect. | |||
Nuetral red or Light green | Counterstain | Ionic bonding | Omitted: Other tissue components will not be demonstrated. |
Too short: Other tissue components will stain weakly. | |||
Too long: Dark counterstain interferes with visualization of reticulin. |
Special Considerations
Ammoniacal silver is reactive and should be nuetralized with table salt or a saline solution before disposal. Use caution.
References
Officer B. HIML251 Lecture notes: Silver Stains. Gordon and Sweet’s Reticulin, March 26, 2009