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, hematoxylin, 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.

73 news items


Wed, 25 Nov 2015 01:38:12 -0800

For histological analysis, the sections were stained for 15 seconds in Mayer's haematoxylin and counterstained for 2 minutes in eosin. Histological severity scoring was performed in a double blinded manner using the following criteria, 0 unaffected ...


Mon, 23 Nov 2015 02:52:30 -0800

Tissues were fixed in either 10% neutral-buffered formalin or 4% paraformaldehyde at 4 °C and embedded in paraffin for sectioning. 5 micron sections were prepared for haematoxylin and eosin staining, immunohistochemistry and immunofluorescence.


Mon, 16 Nov 2015 01:56:15 -0800

The tissue microarray was then stained with a solution of 3, 3-diaminobenzidine tetrahydrochloride and counterstained with haematoxylin. Afterwards, the tissue microarray was assessed by two pathologists who were blinded to clinical and pathologic ...
Tue, 03 Nov 2015 02:53:53 -0800

The SRS microscopy images enabled pathologists to identify normal tissue, infiltrating glioma and dense glioma with the same reliability as they could with haematoxylin and eosin staining. The team also developed a system to automatically quantify the ...


Mon, 09 Nov 2015 03:03:39 -0800

Antibody localization and detection were achieved using Novolink reagents (Leica, Milton Keynes, UK), a diaminibenzidine chromogen and haematoxylin counterstain. Stained TMA sections were microscopically assessed (DGP, GVS) to semi-quantify the ...

Regulatory Focus

Regulatory Focus
Thu, 15 Oct 2015 07:32:01 -0700

Instead of waiting for the disease to progress, sponsors can look for the absence of residual invasive cancer when evaluating haematoxylin and eosin. Guidelines, Comments. EMA to Host Workshop on Extrapolating Efficacy to Discuss Reflection Paper.


Fri, 16 Oct 2015 02:00:00 -0700

The sixth question assessed preferences for treatment of patients who meet the inclusion criteria and the treatment protocol of the Z0011 trial: T1 or T2, clinically node-negative invasive breast cancer ; one or two positive SLNs by routine ...


Tue, 04 Aug 2015 09:23:37 -0700

After MS acquisition removal of the matrix and haematoxylin and eosin (H&E) staining was performed according to standard protocols and optical images were recorded using a MIRAX desk digital slide scanner (Zeiss, Germany). All images, MALDI-MS image ...

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