Masson's trichrome is a three-colour

staining Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the ...

procedure used in

histology Histology, also known as microscopic anatomy or microanatomy, is the branch of biology which studies the microscopic anatomy of biological tissues. Histology is the microscopic counterpart to gross anatomy, which looks at larger structures vis ...

. The recipes evolved from

Claude L. Pierre Masson Claude L. Pierre Masson (November 12, 1880 – May 11, 1959) was a Canadian medical doctor. He was considered one of the leading histopathologists of his era. He was born in Dijon and studied medicine at the University of Paris and the Paste ...

's (1880–1959) original formulation have different specific applications, but all are suited for distinguishing

cells Cell most often refers to: * Cell (biology), the functional basic unit of life Cell may also refer to: Locations * Monastic cell, a small room, hut, or cave in which a religious recluse lives, alternatively the small precursor of a monastery w ...

from surrounding

connective tissue Connective tissue is one of the four primary types of animal tissue, along with epithelial tissue, muscle tissue, and nervous tissue. It develops from the mesenchyme derived from the mesoderm the middle embryonic germ layer. Connective tiss ...

. Most recipes produce red

keratin Keratin () is one of a family of structural fibrous proteins also known as ''scleroproteins''. Alpha-keratin (α-keratin) is a type of keratin found in vertebrates. It is the key structural material making up scales, hair, nails, feathers, ho ...

and

muscle fiber A muscle cell is also known as a myocyte when referring to either a cardiac muscle cell (cardiomyocyte), or a smooth muscle cell as these are both small cells. A skeletal muscle cell is long and threadlike with many nuclei and is called a muscl ...

s, blue or green

collagen Collagen () is the main structural protein in the extracellular matrix found in the body's various connective tissues. As the main component of connective tissue, it is the most abundant protein in mammals, making up from 25% to 35% of the whole ...

and

bone A bone is a Stiffness, rigid Organ (biology), organ that constitutes part of the skeleton in most vertebrate animals. Bones protect the various other organs of the body, produce red blood cell, red and white blood cells, store minerals, provid ...

, light red or pink

cytoplasm In cell biology, the cytoplasm is all of the material within a eukaryotic cell, enclosed by the cell membrane, except for the cell nucleus. The material inside the nucleus and contained within the nuclear membrane is termed the nucleoplasm. The ...

, and dark brown to black

cell nuclei The cell nucleus (pl. nuclei; from Latin or , meaning ''kernel'' or ''seed'') is a membrane-bound organelle found in eukaryotic cells. Eukaryotic cells usually have a single nucleus, but a few cell types, such as mammalian red blood cells, h ...

. The trichrome is applied by immersion of the fixated sample into

Weigert's iron hematoxylin Weigert's elastic stain is a combination of stains used in histology which is useful in identifying elastic fibers. Often orcein or a combination of resorcinol and fuchsine are used for staining. For counterstaining cell nuclei nuclear fast red ...

, and then three different solutions, labeled A, B, and C: * Weigert's

hematoxylin Haematoxylin or hematoxylin (), also called natural black 1 or C.I. 75290, is a compound extracted from heartwood of the logwood tree (''Haematoxylum campechianum'') with a chemical formula of . This naturally derived dye has been used as a h ...

is a sequence of three solutions:

ferric chloride Iron(III) chloride is the inorganic compound with the formula . Also called ferric chloride, it is a common compound of iron in the +3 oxidation state. The anhydrous compound is a crystalline solid with a melting point of 307.6 °C. The colo ...

in diluted

hydrochloric acid Hydrochloric acid, also known as muriatic acid, is an aqueous solution of hydrogen chloride. It is a colorless solution with a distinctive pungent smell. It is classified as a strong acid Acid strength is the tendency of an acid, symbol ...

in 95%

ethanol Ethanol (abbr. EtOH; also called ethyl alcohol, grain alcohol, drinking alcohol, or simply alcohol) is an organic compound. It is an Alcohol (chemistry), alcohol with the chemical formula . Its formula can be also written as or (an ethyl ...

, and

potassium ferricyanide Potassium ferricyanide is the chemical compound with the formula K3 e(CN)6 This bright red salt contains the octahedrally coordinated 3−.html" ;"title="e(CN)6sup>3−">e(CN)6sup>3− ion. It is soluble in water and its solution shows some g ...

solution alkalized by

sodium borate Sodium borate is a generic name for any salt of sodium with an anion consisting of boron and oxygen, and possibly hydrogen, or any hydrate thereof. It can be seen as a hydrated sodium salt of the appropriate boroxy acid, although the latter may not ...

. It is used to stain the nuclei. * Solution A, also called plasma stain, contains

acid fuchsin Acid fuchsin or fuchsine acid, (also called Acid Violet 19 and C.I. 42685) is an acidic magenta dye with the chemical formula C20H17N3Na2O9S3. It is a sodium sulfonate derivative of fuchsine. Acid fuchsin has wide use in histology, and is one of ...

, Xylidine Ponceau,

glacial acetic acid Acetic acid , systematically named ethanoic acid , is an acidic, colourless liquid and organic compound with the chemical formula (also written as , , or ). Vinegar is at least 4% acetic acid by volume, making acetic acid the main component ...

, and

distilled water Distilled water is water that has been boiled into vapor and condensed back into liquid in a separate container. Impurities in the original water that do not boil below or near the boiling point of water remain in the original container. Thus, di ...

. Other red acid dyes can be used, e.g. the

Biebrich scarlet Biebrich scarlet (C.I. 26905) is a molecule used in Lillie's trichrome. The dye was created in 1878 by the German chemist Rudolf Nietzki. Biebrich scarlet dyes are used to color hydrophobic materials like fats and oils. The dye is an illegal d ...

in Lillie's trichrome. * Solution B contains

phosphomolybdic acid Phosphomolybdic acid is the heteropolymetalate with the formula . It is a yellow solid, although even slightly impure samples have a greenish coloration. It is also known as dodeca molybdophosphoric acid or PMA, is a yellow-green chemical compound ...

in distilled water. * Solution C, also called fibre stain, contains

Light Green SF yellowish Light green SF, also called C.I. 42095, light green SF yellowish, is a green triarylmethane dye. Uses Biomedical It is used in histology for staining collagen; for that purpose it is a standard dye in North America. In Masson's trichrome i ...

, or alternatively

Fast Green FCF Fast Green FCF, also called Food green 3, FD&C Green No. 3, Green 1724, Solid Green FCF, and C.I. 42053, is a turquoise triarylmethane food dye. Its E number is E143. Fast Green FCF is recommended as a replacement of Light Green SF yellowish in ...

. It is used to stain collagen. If blue is preferred to green,

methyl blue Methyl blue is a chemical compound with the molecular formula C37H27N3Na2O9S3. It is used as a stain in histology, and stains collagen blue in tissue sections. It can be used in some differential staining techniques such as Mallory's connect ...

water blue Water blue, also known as aniline blue, Acid blue 22, Soluble Blue 3M, Marine Blue V, or C.I. 42755, is a chemical compound used as a stain in histology. Water blue stains collagen blue in tissue sections. It is soluble in water and slightly ...

can be substituted. Standard applications: Masson's trichrome staining is widely used to study muscular pathologies (

muscular dystrophy Muscular dystrophies (MD) are a genetically and clinically heterogeneous group of rare neuromuscular diseases that cause progressive weakness and breakdown of skeletal muscles over time. The disorders differ as to which muscles are primarily affe ...

), cardiac pathologies (

infarct Infarction is tissue death (necrosis) due to Ischemia, inadequate blood supply to the affected area. It may be caused by Thrombosis, artery blockages, rupture, mechanical compression, or vasoconstriction. The resulting lesion is referred to as a ...

), hepatic pathologies (

cirrhosis Cirrhosis, also known as liver cirrhosis or hepatic cirrhosis, and end-stage liver disease, is the impaired liver function caused by the formation of scar tissue known as fibrosis due to damage caused by liver disease. Damage causes tissue repai ...

) or kidney pathologies (glomerular fibrosis). It can also be used to detect and analyze tumors on hepatic and kidney biopsies.

Variants

A common variant is

Lillie's trichrome Lillie's trichrome is a combination of dyes used in histology. It is similar to Masson's trichrome stain, but it uses Biebrich scarlet for the plasma stain. It was initially published by Ralph D. Lillie in 1940. It is applied by submerging the fixa ...

. It is often erroneously called Masson's trichrome. It differs in the dyes used, their concentrations, and the immersion times. Another common variant is the ''Masson trichrome & Verhoeff stain'', which combines the Masson trichrome stain and

Verhoeff's stain Verhoeff's stain, also known as Verhoeff's elastic stain (VEG) or Verhoeff–Van Gieson stain (VVG), is a staining (biology), staining protocol used in histology, developed by United States, American Ophthalmology, ophthalmic surgeon and pathologist ...

. This combination is useful for the examination of

blood vessel The blood vessels are the components of the circulatory system that transport blood throughout the human body. These vessels transport blood cells, nutrients, and oxygen to the tissues of the body. They also take waste and carbon dioxide away ...

s; the Verhoeff stain highlights

elastin Elastin is a protein that in humans is encoded by the ''ELN'' gene. Elastin is a key component of the extracellular matrix in gnathostomes (jawed vertebrates). It is highly elastic and present in connective tissue allowing many tissues in the bod ...

(black) and allows one to easily differentiate small arteries (which typically have at least two elastic laminae) and veins (which have one elastic lamina).

References

External links

Masson's Trichrome Protocol

{{DEFAULTSORT:Masson's Trichrome Stain Staining

Variants

See also

References

External links