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 Sweets Reticulin, March 26, 2009