digplanet beta 1: Athena
Share digplanet:


Applied sciences






















Skeletal formula of haematoxylin
Ball-and-stick model of the haematoxylin molecule
IUPAC name
Other names
Hematoxylin; Natural Black 1; Hematoxyline; Hydroxybrazilin; Hydroxybrasilin; C.I. 75290
517-28-2 YesY
ChemSpider 21106443 YesY
Jmol interactive 3D Image
MeSH Hematoxylin
PubChem 10603
Molar mass 302.28 g·mol−1
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
N verify (what is YesYN ?)
Infobox references

Haematoxylin or hematoxylin (also called 'natural black 1' or 'C.I. 75290') is a compound extracted from the heartwood of the logwood tree.[1] Hematoxylin is a basic / positive compound that binds to and forms salts with acidic, or basophilic, compounds containing negative charges (such as DNA and RNA which are acidic/negative because the nucleic acid building blocks that come off the phosphate backbone are negatively charged) and stains them dark blue or violet. Haematoxylin and eosin together make up haematoxylin and eosin stain, one of the most commonly used stains in histology. This type of stain is a permanent stain as opposed to temporary stains (e.g. iodine solution in KI). Another common stain is phosphotungstic acid haematoxylin, a mix of haematoxylin with phosphotungstic acid. When oxidized it forms haematein, a compound that forms strongly coloured complexes with certain metal ions, the most notable ones being Fe(III) and Al(III) salts. Metal-haematein complexes are used to stain cell nuclei prior to examination under a microscope. Structures that stain with iron- or aluminium-haematein are often called basophilic, even though the mechanism of the staining is different from that of staining with basic dyes.

In the early 1970s and in 2008, there were shortages of haematoxylin due to interruptions in its extraction from logwood. The price of the compound increased, affecting the cost of diagnostic histopathology, and prompted a search for alternative nuclear stains. Before the use of any alternatives became firmly established, haematoxylin returned to the market, though at a higher price, and resumed its place in histopathology. Several synthetic dyes have been recommended as replacements, notably celestine blue (CI 51050), gallocyanine (CI 51030), gallein (CI 45445) and eriochrome cyanine R (also called chromoxane cyanine R and solochrome cyanine (CI 43820). All four have Fe(III) as the mordant. Another alternative is the aluminium complex of oxidized brazilin, which differs from haematoxylin by only one hydroxyl group.

Staining solutions[edit]

Mouse skin showing hematoxylin (nuclear/purple) and eosin (cytosolic/pink).

These stains are commonly employed for histological studies. The mordants used to demonstrate nuclear and cytoplasmic structures are alum and iron, forming lakes or coloured complexes (dye-mordant-tissue complexes), the colour of which will depend on the salt used. Aluminium salt lakes are usually coloured blue-white, whereas ferric salt lakes are coloured blue-black.

Aluminium solutions[edit]

The three main alum haematoxylin solutions employed are Ehrlich's haematoxylin, Harris's haematoxylin, and Mayer's haematoxylin. The name haemalum is preferable to "haematoxylin" for these solutions because haematein, a product of oxidation of haematoxylin, is the compound that combines with aluminium ions to form the active dye-metal complex. Alum haematoxylin solutions impart to the nuclei of cells a light transparent red stain that rapidly turns blue on exposure to any neutral or alkaline liquid.

Alum or potassium aluminium sulfate used as the mordant usually dissociates in an alkaline solution, combining with HO of water to form insoluble aluminium hydroxide. In the presence of excess acid, aluminium hydroxide cannot be formed, thus causing failure of aluminium haematoxylin dye-lake to form, due to lack of OH ions. Hence, acid solutions of alum haematoxylin become red. During staining, alum haematoxylin-stained sections are usually passed on to a neutral or alkaline solution (e.g., hard tap water or 1% ammonium hydroxide) in order to neutralize the acid and form an insoluble blue aluminium haematin complex. This procedure is known as blueing.

When tap water is not sufficiently alkaline, or is even acidic and is unsatisfactory for blueing haematoxylin, a tap water substitute consisting of 3.5 g NaHCO3 and 20 g MgSO4.7H2O in one litre of water with thymol (to inhibit formation of moulds), is used to accelerate blueing of thin paraffin sections. Addition of a trace of any alkali to tap or distilled water also provides an effective blueing solution; a few drops of strong ammonium hydroxide or of saturated aqueous lithium carbonate, added immediately before use, are sufficient for a 400 ml staining dish full of water. Use of very cold water slows down the blueing process, whereas warming accelerates it. In fact, the use of water below 10 °C for blueing sections may even produce pink artifact discolourations in the tissue.

See also[edit]


  1. ^ Cooksey 2010


  • Brown, G. G. (1978). An Introduction to Histotechnology. Appleton-Century-Crofts, New York.
  • Cooksey C. (2010) Hematoxylin and related compounds - an annotated bibliography concerning their origin, properties, chemistry and certain applications. Biotechnic & Histochemistry 85(1): 65-82. PMID 19568968
  • Dapson R., Horobin R.W., Kiernan J.A. (2010) Hematoxylin shortages their causes and duration and other dyes that can replace hemalum in routine hematoxylin and eosin staining. Biotechnic & Histochemistry 85(1): 55-63.
  • Godwin Avwioro (2011). Histochemical Uses Of Haematoxylin - A Review. JPCS 1:24-34. PDF
  • Jocelyn H. Bruce-Gregorios, M.D.: Histopathologic Techniques, JMC Press Inc., Quezon City, Philippines, 1974.
  • Meloan, S. M. & Puchtler, H. 1987. "Harris hematoxylin," what Harris really wrote and the mechanism of hemalum stains. Journal of Histotechnology 10: 257-261.
  • Puchtler, H., Meloan, S.N. & Waldrop, F.S. 1986. Application of current chemical concepts to metal-haematein and -brazilein stains. Histochemistry 85: 353-364.

External links[edit]

Original courtesy of Wikipedia: http://en.wikipedia.org/wiki/Haematoxylin — Please support Wikipedia.
This page uses Creative Commons Licensed content from Wikipedia. A portion of the proceeds from advertising on Digplanet goes to supporting Wikipedia.

124 news items


Wed, 03 Feb 2016 10:16:54 -0800

a, Expression (in situ hybridization) of Lmx1a, Eomes, Lhx2, and Atoh1 in the embryonic cerebellum (e13.5). b, Immunofluorescence microscopy for the TFs shown in a performed on sagittal sections of the e13.5 murine cerebellum. c, Haematoxylin and ...


Fri, 05 Feb 2016 07:33:45 -0800

Formalin-fixed samples were embedded in paraffin (FFPE) after dehydration in ascending alcohol concentrations ending in xylol, with 3 μm sections mounted on positively charged glass slides and used for routine haematoxylin and eosin (HE) staining, ...


Tue, 19 Jan 2016 01:28:30 -0800

However, the tendons were evaluated by using a classic tendon histology score, in which the tissue samples were stained with haematoxylin and eosin and alcian blue. This histology score has been previously used to evaluate a similar rabbit model of ...


Wed, 13 Jan 2016 05:11:46 -0800

The fourth section from each block was cut with a thickness of 3 μm and stained with haematoxylin and eosin to identify tumour-enriched regions. This section was used as a template for macrodissection of the tumour-enriched region from the remaining ...


Tue, 10 Nov 2015 08:18:45 -0800

The SH-SY5Y tumours were harvested from mice and then sectioned and stained with haematoxylin and eosin (H&E) to observe histopathological changes (Fig. 8c–g). Xenografts treated with GB1–biosilica (Fig. 8d) or anti-p75NTR–GB1–biosilica (Fig.


Tue, 01 Dec 2015 04:35:00 -0800

Haematoxylin-eosin staining was carried out by cutting 2 mm sample slices, which were mounted and stained on poly-lysine slides. Samples were observed by light microscopy using a Nikon i55 (Nikon Corporation, Tokyo, Japan) optical microscope.


Wed, 09 Sep 2015 09:56:27 -0700

Tissue samples were immersed in 10% buffered formalin and prion infectivity was inactivated by immersion into 98% formic acid for one hour. Tissue samples were processed to paraffin wax and tissue sections were routinely stained with haematoxylin and ...


Fri, 18 Dec 2015 02:30:00 -0800

Phosphotungstic acid haematoxylin–stained sections showed that collagen fibres were deposited and arranged irregularly in the caval wall and thrombus at 14 days (Fig. 4A,B). Recanalization channels were also observed at the margins of the thrombus (Fig.

Oops, we seem to be having trouble contacting Twitter

Support Wikipedia

A portion of the proceeds from advertising on Digplanet goes to supporting Wikipedia. Please add your support for Wikipedia!

Searchlight Group

Digplanet also receives support from Searchlight Group. Visit Searchlight