Histology Stains

From Embryology

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 [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 - pinkmuscle - 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

Hematoxylin and Eosin

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.

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.


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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