A technique is described for the isolation of colonies of flax rust (Melampsora lini (Ehrenb.) Lév., Race No. 3) from infected cotyledons. The technique depends on the digestion of the host cell walls with hydrolytic enzymes and washing of the liberated colonies. It is thus possible to collect large numbers of flax-rust colonies with only a few host cells adhering to them. Axenic cultures were established from colonies isolated in this way. This technique may be useful in establishing axenic cultures of other rust fungi.