H&E: The Bread and Butter Stain

The Journal of Histotechnology, NSH’s peer reviewed histology Journal, has added a new section to their website, Most Read Journal Articles. The page features the most popular articles from the Journal for the past two years, and has made them open access, so both NSH members and non-members alike can read them.

So what are histology Journal readers most interested in these days? Well… stains! Obviously. But one stain in particular dominates two of the leading articles and is mentioned in several other editorials; the Hematoxylin and Eosin stain, commonly referred to as H&E.

Stomach biopsy stained with Gill’s B hematoxylin and eosin–phloxine, ×40. Mucin is unstained.- Tony Henwood

Tony Henwood, author of one of the featured articles, refers to the H&E stain as “the bread and butter stain in histopathology”. H&E is known for being the basis of histology staining knowledge, and is one of the oldest staining combinations used. Haematoxylin as a dye dates back as early as the 1500’s, obtained from logwood trees by the Spanish. It was soon traded as a fabric dye, and introduced into scientific staining in the mid-1800’s through a series of discoveries by scientists such as Bohmer in 1865 who added alum, and Ehrlich in 1886 who added acetic acid to the solution.

Despite its age and relative simplicity, the H&E stain remains one of the most commonly used stains in modern histology. Clearly, there is still much to discover about it as well!

Check out these top journal articles featuring the ever-popular H&E.

“Hematoxylin and eosin staining of mucins of the gastrointestinal tract” by Anthony Henwood

An infrequent observation of assessing hematoxylin and eosin sections is the blue staining of mucins (for example those in goblet cells). This is believed to be due to a low concentration of alum and high pH of the hematoxylin staining solution. This study examines the incidence of blue mucin in various sites of the gastrointestinal tract using a low alum, high pH hematoxylin solution.

“A combination Prussian blue – hematoxylin and eosin staining technique for identification of iron and other histological features” by Katelyn Rowatt, Rachel E. Burns, Salvatore Frasca Jr. & Denise M. Long

The Prussian blue reaction (PB) detects ferric iron in histological sections but the nuclear fast red (NFR) counterstain does not selectively stain the surrounding tissue and cellular features very well. The PB/NFR stain has the advantage of detecting iron located in tissue sections, but a significant disadvantage of having poorly differentiated tissue components, as compared to a routine hematoxylin and eosin (H&E). We developed a combination of Gomori’s Prussian blue/H&E staining method (PB/H&E), and modified the technique for best performance and clarity, then assessed the ability of this new combination stain to differentiate histological features of the tissue and identify iron.

1. Anthony F. Henwood (2017) Hematoxylin and eosin staining of mucins of the gastrointestinal tract, Journal of Histotechnology, 40:1, 21-24, DOI: 10.1080/01478885.2017.1264556

2. M Titford (2005) The long history of hematoxylin, Biotechnic & Histochemistry, 80:2, 73-78, DOI: 10.1080/10520290500138372

3. https://jcp.bmj.com/content/jclinpath/50/9/716.full.pdf

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The National Society for Histotechnology is a professional member organization for individuals actively involved in the histology profession. NSH has over 3,000 members worldwide, and is the leading provider of histology focused continuing education.  



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