Eosin

Last updated
Eosin Y Eosin Y Structural Formulae V.1.svg
Eosin Y
Eosin B Eosin B Structural Formulae V.1.svg
Eosin B

Eosin is the name of several fluorescent acidic compounds which bind to and form salts with basic, or eosinophilic, compounds like proteins containing amino acid residues such as arginine and lysine, and stains them dark red or pink as a result of the actions of bromine on eosin. In addition to staining proteins in the cytoplasm, it can be used to stain collagen and muscle fibers for examination under the microscope. Structures that stain readily with eosin are termed eosinophilic. In the field of histology, Eosin Y is the form of eosin used most often as a histologic stain. [1] [2]

Contents

Etymology

Eosin was named by its inventor Heinrich Caro after the nickname (Eos) of a childhood friend, Anna Peters. [3]

Variants

Eosin Y solution for staining microscopy slides Eosin solution.jpg
Eosin Y solution for staining microscopy slides

There are actually two very closely related compounds commonly referred to as eosin. Most often used is in histology is Eosin Y [1] [2] (also known as eosin Y ws, eosin yellowish, Acid Red 87, C.I. 45380, bromoeosine, bromofluoresceic acid, D&C Red No. 22); it has a very slightly yellowish cast. The other eosin compound is Eosin B (eosin bluish, Acid Red 91, C.I. 45400, Saffrosine, Eosin Scarlet, or imperial red); it has a very faint bluish cast. The two dyes are interchangeable, and the use of one or the other is a matter of preference and tradition.

Eosin Y is a tetrabromo derivative of fluorescein. [4] Eosin B is a dibromo dinitro derivative of fluorescein. [5]

Uses

Use in histology

Eosinophilic staining, compared to other patterns when using hematoxylin and eosin (H&E) Eosinophilic, basophilic, chromophobic and amphophilic staining.png
Eosinophilic staining, compared to other patterns when using hematoxylin and eosin (H&E)

Eosin is most often used as a counterstain to hematoxylin in H&E (haematoxylin and eosin) staining. H&E staining is one of the most commonly used techniques in histology. Tissue stained with haematoxylin and eosin shows cytoplasm stained pink-orange and nuclei stained darkly, either blue or purple. Eosin also stains red blood cells intensely red.

For staining, eosin Y is typically used in concentrations of 1 to 5 percent weight by volume, dissolved in water or ethanol. [6] For prevention of mold growth in aqueous solutions, thymol is sometimes added. [7] A small concentration (0.5 percent) of acetic acid usually gives a deeper red stain to the tissue.

It is listed as an IARC class 3 carcinogen.

Other uses

In his Field with Irises near Arles painting, Van Gogh incorporated red eosin dye into the color of the irises depicted at the bottom of the painting. Due to eosin's tendency to fade, the petals have now attained a blueish hue from their original purple coloration. Veld met irissen bij Arles - s0037V1962 - Van Gogh Museum.jpg
In his Field with Irises near Arles painting, Van Gogh incorporated red eosin dye into the color of the irises depicted at the bottom of the painting. Due to eosin's tendency to fade, the petals have now attained a blueish hue from their original purple coloration.

Eosin is also used as a red dye in inks; however, the molecule, especially that of eosin Y, tends to degrade over time, leaving behind its bromine atoms, hence causing paint incorporating such a dye to obtain a darker brown tinge over time. [8] A notable user of eosin dye was the post-impressionist painter Van Gogh.

See also

Related Research Articles

<span class="mw-page-title-main">Haematoxylin</span> Natural stain derived from hearthwood and used in histology

Haematoxylin or hematoxylin, also called natural black 1 or C.I. 75290, is a compound extracted from heartwood of the logwood tree with a chemical formula of C
16
H
14
O
6
. This naturally derived dye has been used as a histologic stain, ink and as a dye in the textile and leather industry. As a dye, haematoxylin has been called Palo de Campeche, logwood extract, bluewood and blackwood. In histology, haematoxylin staining is commonly followed (counterstained), with eosin, when paired, this staining procedure is known as H&E staining, and is one of the most commonly used combinations in histology. In addition to its use in the H&E stain, haematoxylin is also a component of the Papanicolaou stain which is widely used in the study of cytology specimens.

<span class="mw-page-title-main">Mordant</span> Substance used for binding dyes to fabrics

A mordant or dye fixative is a substance used to set dyes on fabrics by forming a coordination complex with the dye, which then attaches to the fabric. It may be used for dyeing fabrics or for intensifying stains in cell or tissue preparations. Although mordants are still used, especially by small batch dyers, it has been largely displaced in industry by directs.

<span class="mw-page-title-main">Staining</span> Technique used to enhance visual contrast of specimens observed under a microscope

Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology, in cytology, and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues, cell populations, or organelles within individual cells.

<span class="mw-page-title-main">Fluorescein</span> Synthetic organic compound used as dye and fluorescent tracer

Fluorescein is an organic compound and dye based on the xanthene tricyclic structural motif, formally belonging to triarylmethine dyes family. It is available as a dark orange/red powder slightly soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications.

<span class="mw-page-title-main">Eosinophilic</span> Quality of being colored by eosin

Eosinophilic is the staining of tissues, cells, or organelles after they have been washed with eosin, a dye.

<span class="mw-page-title-main">Basophilic</span>

Basophilic is a technical term used by pathologists. It describes the appearance of cells, tissues and cellular structures as seen through the microscope after a histological section has been stained with a basic dye. The most common such dye is haematoxylin.

<span class="mw-page-title-main">Methyl blue</span> Chemical compound

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 connective tissue stain and Gömöri trichrome stain, and can be used to mediate electron transfer in microbial fuel cells. Fungal cell walls are also stained by methyl blue.

<span class="mw-page-title-main">Eosin Y</span> Chemical compound

Eosin Y, also called C.I. 45380 or C.I. Acid Red 87, is a member of the triarylmethane dyes. It is produced from fluorescein by bromination.

<span class="mw-page-title-main">Light green SF</span> Chemical compound

Light green SF, also called C.I. 42095, light green SF yellowish, is a green triarylmethane dye.

<span class="mw-page-title-main">Papanicolaou stain</span> Histological staining method

Papanicolaou stain is a multichromatic (multicolored) cytological staining technique developed by George Papanicolaou in 1942. The Papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. Although most notable for its use in the detection of cervical cancer in the Pap test or Pap smear, it is also used to stain non-gynecological specimen preparations from a variety of bodily secretions and from small needle biopsies of organs and tissues. Papanicolaou published three formulations of this stain in 1942, 1954, and 1960.

<span class="mw-page-title-main">H&E stain</span> Histological stain method

Hematoxylin and eosin stain is one of the principal tissue stains used in histology. It is the most widely used stain in medical diagnosis and is often the gold standard. For example, when a pathologist looks at a biopsy of a suspected cancer, the histological section is likely to be stained with H&E.

<span class="mw-page-title-main">Phosphotungstic acid-haematoxylin stain</span> Biological stain used for staining of tissues

Phosphotungstic acid haematoxylin (PTAH) is a mix of haematoxylin with phosphotungstic acid, used in histology for staining.

<span class="mw-page-title-main">Phosphotungstic acid</span> Chemical compound

Phosphotungstic acid (PTA) or tungstophosphoric acid (TPA), is a heteropoly acid with the chemical formula H3PW12O40]. It forms hydrates H3[PW12O40nH2O. It is normally isolated as the n = 24 hydrate but can be desiccated to the hexahydrate (n = 6). EPTA is the name of ethanolic phosphotungstic acid, its alcohol solution used in biology. It has the appearance of small, colorless-grayish or slightly yellow-green crystals, with melting point 89 °C (24 H2O hydrate). It is odorless and soluble in water (200 g/100 ml). It is not especially toxic, but is a mild acidic irritant. The compound is known by a variety of names and acronyms (see 'other names' section of infobox).

<span class="mw-page-title-main">Xanthene</span> Chemical compound used to make dyes

Xanthene (9H-xanthene, 10H-9-oxaanthracene) is the organic compound with the formula CH2[C6H4]2O. It is a yellow solid that is soluble in common organic solvents. Xanthene itself is an obscure compound, but many of its derivatives are useful dyes.

<span class="mw-page-title-main">Acridine orange</span> Organic dye used in biochemistry

Acridine orange is an organic compound that serves as a nucleic acid-selective fluorescent dye with cationic properties useful for cell cycle determination. Acridine orange is cell-permeable, which allows the dye to interact with DNA by intercalation, or RNA via electrostatic attractions. When bound to DNA, acridine orange is very similar spectrally to an organic compound known as fluorescein. Acridine orange and fluorescein have a maximum excitation at 502nm and 525 nm (green). When acridine orange associates with RNA, the fluorescent dye experiences a maximum excitation shift from 525 nm (green) to 460 nm (blue). The shift in maximum excitation also produces a maximum emission of 650 nm (red). Acridine orange is able to withstand low pH environments, allowing the fluorescent dye to penetrate acidic organelles such as lysosomes and phagolysosomes that are membrane-bound organelles essential for acid hydrolysis or for producing products of phagocytosis of apoptotic cells. Acridine orange is used in epifluorescence microscopy and flow cytometry. The ability to penetrate the cell membranes of acidic organelles and cationic properties of acridine orange allows the dye to differentiate between various types of cells. The shift in maximum excitation and emission wavelengths provides a foundation to predict the wavelength at which the cells will stain.

<span class="mw-page-title-main">Acidophile (histology)</span>

Acidophile is a term used by histologists to describe a particular staining pattern of cells and tissues when using haematoxylin and eosin stains. Specifically, the name refers to structures which "love" acid, and take it up readily. More specifically, acidophilia can be described by cationic groups of most often proteins in the cell readily reacting with acidic stains.

<span class="mw-page-title-main">Acid fuchsin</span> Chemical compound

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 the dyes used in Masson's trichrome stain. This method is commonly used to stain cytoplasm and nuclei of tissue sections in the histology laboratory in order to distinguish muscle from collagen. The muscle stains red with the acid fuchsin, and the collagen is stained green or blue with Light Green SF yellowish or methyl blue. It can also be used to identify growing bacteria.

Infantile digital fibromatosis (IDF), also termed inclusion body fibromatosis, Reye tumor, or Reye's tumor, usually occurs as a single, small, asymptomatic, nodule in the dermis on a finger or toe of infants and young children. IMF is a rare disorder with approximately 200 cases reported in the medical literature as of 2021. The World Health Organization, 2020, classified these nodules as a specific benign tumor type in the category of fibroblastic and myofibroblastic tumors. IDF was first described by the Australian pathologist, Douglas Reye, in 1965.

Trichrome stains are staining methods in which three anionic dyes are used, in conjunction with either phosphomolybdic acid (PMA), phosphotungstic acid (PTA), or a mixture of these heteropolyacids. Probably the first trichrome method was that of Frank B Mallory, an American pathologist, first published in 1900. Unfortunately, none of Mallory's publications provide any explanation of the rationales of either his trichrome or his phosphotungstic acid-haematoxylin (PTAH) method. Nobody knows why Mallory introduced heteropolyacids into microtechnique.

<span class="mw-page-title-main">Red neuron</span>

A "red neuron" is a pathological finding in neurons, generally of the central nervous system, indicative of acute neuronal injury and subsequent apoptosis or necrosis. Acidophilic neurons are often found in the first 12–24 hours after an ischemic injury such as a stroke. Since neurons are permanent cells, they are most susceptible to hypoxic injury. The red coloration is due to pyknosis or degradation of the nucleus and loss of Nissl bodies which are normally stained blue (basophilic) on hematoxylin & eosin staining. This leaves only the degraded proteins which stains red (eosinophilic). Acidophilic neurons also can be stained with acidic dyes other than eosin.

References

  1. 1 2 Lillie, Ralph Dougall (1977). H. J. Conn's Biological stains (9th ed.). Baltimore: Williams & Wilkins. pp. 692p.
  2. 1 2 Bancroft, John; Stevens, Alan, eds. (1982). The Theory and Practice of Histological Techniques (2nd ed.). Longman Group Limited.
  3. Travis, Anthony S (1998). ""Ambitious and Glory Hunting . . . Impractical and Fantastic": Heinrich Caro at BASF". Technology and Culture. 39 (1): 105–115. doi:10.2307/3107005. JSTOR   3107005.
  4. Its CAS number is 17372-87-1 and its SMILES structure is O=C5C(Br)=C2O C1=C(Br)C([O-]) =C(Br)C=C1C(C4=C (C([O-])=O)C=C C=C4)=C2C=C3Br.
  5. Its CAS number is 548-28-3 and its SMILES structure is O=C5C(Br)=C2O C1=C(Br)C([O-]) =C([N+]([O-])=O) C=C1C(C4=C(C([O-]) =O)C=CC=C4)=C2 C=C3[N+]([O-])=O.
  6. "Haematoxylin Eosin (H&E) staining". protocolsonline.com. 11 April 2010. Retrieved 22 April 2018.
  7. Hitokoto H, Morozumi S, Wauke T, Sakai S, Kurata H (1980). "Inhibitory effects of spices on growth and toxin production of toxigenic fungi". Appl. Environ. Microbiol. 39 (4): 818–22. doi:10.1128/AEM.39.4.818-822.1980. PMC   291425 . PMID   6769391.
  8. 1 2 "Van Gogh's Fading Colors Inspire Scientific Inquiry".