Histology stains

From CellBiology

Introduction

This page gives a general overview of some histological stains used to identify structures in cells and tissues. This stains information should also be considered in relation to Histology fixatives.

Medicine Foundations students do not need to know this detail.


Common Stains and Their Reactions

Stain
Nucleus
Cytoplasm
Collagen
RBCs
Other
Haematoxylin
blue
-
-
-
mucins - light blue
Eosin
-
pink
pale pink
bright red
colloid - pink muscle - red
IronHaematoxylin
blue/black
-
-
-
Van Gieson
-
brown/yellow
red
yellow
muscle: yellow/browncartilage - pink
Verhoeff's Elastin
black
-
-
-
elastic fibres - black
Tartrazine
-
yellow
yellow
yellow
Silver Impregnation
-
-
grey/brown
-
reticular fibres - black
Methyl Green
dark green
light green
light green
green
Nuclear Fast Red
red
pink
pink
pink
Gomori's Trichrome
purple/red
purple
green
red
keratin - redmuscle - purple/red
Heidenhain's Azan
red
purple/red
deep blue
red
muscle - red
Osmium tetroxide
-
-
brown
brown
myelin, lipids - black
Alcian Blue
-
-
-
-
mucins, - blue
Periodic acid-Schiff's reagent (PAS)
-
-
pink
-
mucins, glycogen, glycocalyx - magenta
PTAH
blue
-
red
blue
muscle bands - blue
Masson's Trichrome
blue/black
red
green/blue
red
cartilage, mucins - blue or green; muscle - red
Luxol Fast Blue
-
-
-
variable
myelin - blue~
Aldehyde Fuchsin
-
-
-
-
elastic fibres, mast cells - deep purple
Light Green
-
-
light green
-
Gallocyanin
dark blue
-
-
-
nucleic acids, Nissl granules - dark blue
Romanowsky(e.g. Leishman's stain)
blue
pink
acidophils - redbasophils - blueazurophilic - purple
Aldehyde Pararosanilin elastic fibres - purple

Supporting Stain Information

Eosin

  • Stains cytoplasm pink to red; red blood cells are also bright red.
  • Common counterstain to hematoxylin.
  • Stain intensity varies with the formula as well as the fixative.

Hematoxylin

  • Stains nuclei blue to dark-blue.
  • Stains the matrix of hyaline cartilage, myxomatous, and mucoid material pale blue.
  • Stains myelin weakly but is not noticeable if combined with eosin stain.

Periodic acid-Schiff (PAS)

  • Stains glycogen, mucin, fungus, basement membrane and other substances.
  • Stain used to detect fungal organisms and cytoplasmic accumulation of glycogen.
  • Stains lysosomes granules red-purple, can be used in recognition of macrophages.

Alcian Blue

  • Stains mucopolysaccharides or glycosaminoglycans
  • cationic dye (positively charged molecule) for the demonstration of glycosaminoglycans.
  • binds anionic (negative) sites on the polysaccharide.
  • can be combined with H&E and VG staining methods.

Masson’s Trichrome Stain

  • Stains nuclei deep blue, skeletal and smooth muscles red, collagen and mucin blue.
  • Stains brain and spinal cord parenchymal tissue dusky pink to red.
  • Used to evaluate fibrosis
  • Striations in skeletal muscles also shows up much better in Masson’s trichrome than in hematoxylin and eosin stain.
    • Although called a trichrome, four dyes (hematoxylin, Biebrich scarlet, acid fuchsin, and analine blue) are utilized.

PhosphoTungstic Acid Hematoxylin (PTAH)

  • Stains nucleus and cytoplasm detail and connective tissue fibers.
  • Stains collagen pink, fibrin blue, and striated muscle blue.
  • Historic stain used to show CNS reactive astrocytes now used immunochemistry for glial fibrillary acidic protein (GFAP).

Toulidine Blue

  • Stains nucleus blue and cytoplasm light blue.
  • A synthetic dye in the thiazins family.

Verhoeff-Van Gieson

  • Verhoeff-Van Gieson or elastic-Van Gieson (EVG) stain.
  • This is a combination of Verhoeff’s elastic stain which is a hematoxylin stain containing ferric chloride and Wright’s iodine solution and Van Gieson stain which contains acid fuchsin, picric acid, and hematoxylin.
  • Stains elastic fibers blue-black to black, collagen pale red, other tissue elements yellow, and nuclei blue to black.


Some text modified from: Theory and practice of histological techniques. 3rd edt. By Bancroft JD and Stevens A. Churchill Livingstone, 1990.


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