Trichrome staining is a process that involves staining combinations that usually contain three dyes of contrasting colours selected to stain connective tissue, muscle, cytoplasm and nuclei in bright colours.
The English pathologist Heneage Gibbes invented the trichrome staining process in 1880 after experimenting with various histological stains. Here is the trichrome staining (which he called “treble staining”) method he describes in his 1880 publication:
- He first stained the histologic sections with picrocarmine and soaked them in acidulated water.
- He then took a roseine solution and diluted it with alcohol and then immersed the same sections in the solution for 2-3 minutes.
- Then, he removed the sections and poured denatured alcohol on them to wash off the excess dye.
- He then placed the sections in a diluted solution of iodine green, after which they were ready for viewing once removed.
On the application of this “treble stain” to histologic sections, he states, “This staining process is well shown in a section of the base of a cat or dog’s tongue, cut through one of the circumvallate papillae, the section should be sufficiently large to include some of the mucous glands, of which there are a large number in that region. If the staining is well done it will show all the muscle fibres stained with picro-carmine, the connective tissue, protoplasm of cells, &c., stained with rosein; while all the nuclei in the superficial epithelium, serous glands, non-striped muscle tissue in the vessels, and elsewhere, are stained a brilliant green.” Here is a photomicrograph of a section of a rat’s airway stained with Masson’s trichrome (nuclei are stained dark red/purple, cytoplasm is stained red/pink and connective tissue is stained blue):
[ux_image_box img=”1547″ image_width=”60″ link=”https://en.wikipedia.org/wiki/Masson%27s_trichrome_stain” target=”_blank”]
Here is a photograph of Heneage Gibbes:
[ux_image_box img=”1550″ image_width=”30″ link=”https://kingscollections.org/victorianlives/g-i/heneage-gibbes-heneage” target=”_blank”]