Mnemonic

Common Histological Stains

A memory aid for the common stains and what each demonstrates.

Expansion

H and E for routine work, with special stains for specific components

Mnemonic

Match the stain to what it demonstrates:

  • Haematoxylin and eosin - the default. Haematoxylin stains nuclei blue (basophilic), eosin stains cytoplasm pink (eosinophilic)
  • Periodic acid-Schiff (PAS) - glycogen, glycoproteins, basement membranes, fungi. Magenta
  • Congo red - amyloid, giving apple green birefringence under polarised light
  • Ziehl-Neelsen - acid fast bacilli, so mycobacteria. Red on blue
  • Prussian blue (Perls) - iron, for haemochromatosis
  • Masson’s trichrome - collagen and fibrosis, blue or green
  • Gram - bacteria, purple positive and pink negative
  • Silver stains - reticulin, fungi (Grocott), neurofibrillary tangles
  • Oil red O and Sudan - lipid, requiring frozen sections

“H&E for everything, PAS for sugar, Congo red for amyloid, ZN for TB, Perls for iron.”

Apple green birefringence with Congo red is diagnostic of amyloid and is the single most frequently examined stain result.

Expansion

Stain Demonstrates Appearance
Haematoxylin and eosin Routine Nuclei blue, cytoplasm and collagen pink
PAS Glycogen, glycoprotein, basement membrane, fungi Magenta
PAS with diastase Distinguishes glycogen (digested) from other PAS-positive material
Congo red Amyloid Apple green birefringence under polarised light
Ziehl-Neelsen Acid fast bacilli (mycobacteria) Red on blue
Gram Bacteria Gram positive purple, negative pink
Masson trichrome Collagen and fibrosis Collagen blue or green
Perls Prussian blue Iron (haemosiderin) Blue
Oil red O, Sudan Lipid; needs frozen section Red
Silver stains Reticulin, fungi, spirochaetes, neurofibrillary tangles Black
Alcian blue Acid mucins Blue
Toluidine blue Mast cell granules (metachromasia) Purple

Basophilic structures (nuclei, ribosomes, rough endoplasmic reticulum) bind the basic dye haematoxylin and appear blue; eosinophilic (acidophilic) structures such as cytoplasmic protein, collagen and red cells appear pink.

Congo red with apple green birefringence remains the definitive test for amyloid, and requires polarised light.