Ink and Vinegar, a Simple Staining Technique for Arbuscular-Mycorrhizal Fungi

Developed by Horst Vierheilig

Estacion Experimental del Zaidin, Consejo Superior de Investigaciones Cientificas, Calle Prof. Albareda 1, Apdo. 419, E-18008 Granada, Spain. (Current address)

in collaboration with: Andrew P. Coughlan, and Yves Piche - Laval University, Quebec, Canada and Urs Wyss- Kiel University, Germany

Hyphae and arbuscule in
cucumber roots

Hyphae and vesicles in roots
of Idria columnaris

Hyphae and arbuscule in roots of
Idria columnaris, an ancient tree
from the desert of Baja
California, Mexico

A reliable, inexpensive, and simple method for staining arbuscular-mycorrhizal fungal colonizations in root tissues was developed. Apart from applications in research, this nontoxic, high-quality staining method also could be of great utility in teaching exercises. After adequate clearing with KOH, an ink-vinegar solution successfully stained all fungal structures, rendering them clearly visible.

This technique was developed in order to substitute conventional staining techniques of mycorrhizal fungi using potentially carcinogenic dyes by substituting them with harmless inks.

This website containstechnical specifications of the procedure and tips, a list of necessary chemicals, photographs (in the photo gallery), and a PDF file of the publication that originally describes the entire procedure and research paper using this technique.