A modified procedure for staining arbuscular mycorrhizal fungi in roots

Roots were cleared by boiling in 10% KOH and rinsed several times with tap water. Thereafter roots were boiled in the staining solutions. The staining solutions consisted in different dyes (trypban blue (0.05%), anilin blue (0.05%) and acid fuchsin (0.01%)) which were dissolved in usual household vinegar (5% acetic acid). For best contrast, tryphan and anilin blue stained roots were destained with tap water, whereas acid fuchsin stained roots were destained with vinegar. All fungal structures were stained and clearly visible.