Verhoeff's stain, also known as Verhoeff's elastic stain (VEG) or Verhoeff–Van Gieson stain (VVG), is a

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

protocol used in

histology Histology, also known as microscopic anatomy or microanatomy, is the branch of biology that studies the microscopic anatomy of biological tissue (biology), tissues. Histology is the microscopic counterpart to gross anatomy, which looks at large ...

, developed by American ophthalmic surgeon and pathologist Frederick Herman Verhoeff (1874–1968) in 1908. The formulation is used to demonstrate normal or pathologic

elastic fiber Elastic fibers (or yellow fibers) are an essential component of the extracellular matrix composed of bundles of proteins (elastin) which are produced by a number of different cell types including fibroblasts, endothelial, smooth muscle, and ai ...

s. Verhoeff's stain forms a variety of cationic, anionic and non-ionic bonds with

elastin Elastin is a protein encoded by the ''ELN'' gene in humans and several other animals. Elastin is a key component in the extracellular matrix of gnathostomes (jawed vertebrates). It is highly Elasticity (physics), elastic and present in connective ...

, the main constituent of elastic fiber tissue. Elastin has a strong affinity for the iron-hematoxylin

complex Complex commonly refers to: * Complexity, the behaviour of a system whose components interact in multiple ways so possible interactions are difficult to describe ** Complex system, a system composed of many components which may interact with each ...

formed by the reagents in the stain and will hence retain

dye Juan de Guillebon, better known by his stage name DyE, is a French musician. He is known for the music video of the single "Fantasy Fantasy is a genre of speculative fiction that involves supernatural or Magic (supernatural), magical ele ...

longer than other tissue elements. This allows elastin to remain stained while the remaining tissue elements are decolorized.

Sodium thiosulfate Sodium thiosulfate (sodium thiosulphate) is an inorganic compound with the formula . Typically it is available as the white or colorless pentahydrate (x = 5), which is a white solid that dissolves well in water. The compound is a reducing agent an ...

is used to remove excess iodine and a counterstain (most often Van Gieson's stain) is used to contrast the principal stain. Elastic fibers and

cell nuclei The cell nucleus (; : nuclei) 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, have no nuclei, and a few others including osteoc ...

are stained black,

collagen fibers Type I collagen is the most abundant collagen of the human body, consisting of around 90% of the body's total collagen in vertebrates. Due to this, it is also the most abundant protein type found in all vertebrates. Type I forms large, eosinoph ...

are stained red, and other tissue elements including

cytoplasm The cytoplasm describes all the material within a eukaryotic or prokaryotic cell, enclosed by the cell membrane, including the organelles and excluding the nucleus in eukaryotic cells. The material inside the nucleus of a eukaryotic cell a ...

are stained yellow.

Reagents

Verhoeff's stain is a combination of the following reagents: *

Haematoxylin Haematoxylin American and British English spelling differences#ae and oe, or hematoxylin (), also called natural black 1 or Colour Index International, C.I. 75290, is a chemical compound, compound extracted from wood#Heartwood and sapwood, heart ...

Iron(III) chloride Iron(III) chloride describes the inorganic compounds with the formula (H2O)x. Also called ferric chloride, these compounds are some of the most important and commonplace compounds of iron. They are available both in anhydrous and in hydrated f ...

* Lugol's iodine * Van Gieson's stain * Acid fuchsin *

Picric acid Picric acid is an organic compound with the formula (O2N)3C6H2OH. Its IUPAC name is 2,4,6-trinitrophenol (TNP). The name "picric" comes from (''pikros''), meaning "bitter", due to its bitter taste. It is one of the most acidic phenols. Like ot ...

Variants

A common variant is the ''Masson trichrome and Verhoeff stain'', which combines the Masson's trichrome stain and Verhoeff stain. It is sometimes just referred to as a ''Masson trichrome''. This combination is useful for the examination of

blood vessel Blood vessels are the tubular structures of a circulatory system that transport blood throughout many Animal, animals’ bodies. Blood vessels transport blood cells, nutrients, and oxygen to most of the Tissue (biology), tissues of a Body (bi ...

s; the Verhoeff stain highlights

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

References

{{Commonscat, Verhoeff’s stain Staining