Fixation on Histology

Art of the Stain 2021 Contest Winner

Christine Carreira is a Research Assistant working in the WHO/IARC Classification of Tumours Group (WCT) at the International Agency for Research on Cancer IARC part of the World Health Organization. Christine was the winner of the Histology Professionals Day 2021 Art of the Stain Contest. In this post, she shares some background on this beautiful stain.

Art of the Stain Contest 2021

Utility of the Stain: Toluidine Blue/Alcian Yellow Stain is one of the most popular non-silver methods for screening and detection of Helicobacter pylori in gastrointestinal biopsy specimens, where bacteria are stained blue in contrast to yellow mucins. It is a good alternative to Giemsa staining.

Helicobacter pylori (Hp) gastritis is a worldwide problem significantly associated with duodenal and gastric peptic ulcer disease, gastric carcinomas, and MALT-type lymphomas.

This stain was submitted as part of a project (ENIGMA) led by the Prevention and Implementation (PRI) Group at IARC which aims to better understanding the etiology and epidemiology of gastric carcinomas (GC) and Hp infection, and evaluates different strategies for GC prevention.

Principles of Staining: In the first step Alcian Yellow is specifically bound for previously oxidized mucins by periodic acid; they form an insoluble compound that will not be affected by the second part of the procedure when toluidine blue solution will stain gastric bacteria, as well as cellular nuclei and other structures using Toluidine Blue.

This provides excellent contrast between dark blue stained bacteria placed on a yellow surface and extracellular foveolar mucins.

The staining was carried out manually using the Alcian Yellow - Toluidine Blue kit from Diapath (kit code 010269).


Staining is very much an art and Histology is not only a professional field of study but an art form. 

Every work of art hides a story within the patterns of its canvas and every histological image tells a story of a function or disease.

In the same state of mind as the 2019 winner we can compare this stain to that of a painting by Vincent Van Gogh.

Starry Night Over the Rhône, by van Gogh, was painted on the bank of the Rhône which is the river that crosses our pretty city of Lyon, FRANCE where the IARC (International Agency for Research on Cancer) is located.

Art of the Stain Contest 2021