An efficient protocol was developed for Micropropagation of common ash (Fraxinus excelsior L.). The in vitro culture was established from shoots of 60 years old plus trees. Single, two and three node segments of decapitated shoots were cultured on MS medium supplemented with 1, 2, or 4 mg L1 BAP and 0.1 mg L1 IAA for induction of axillary shoots. The best multiplication rate (7.6) was achieved using single node explants on medium with 1 and 2 mg L1 BAP. Shoots on all the explant types were significantly taller on the medium with 2 mg L1 BAP. Efficiency of axillary shoot formation was the highest in distal nodes and reduced toward proximal nodes of explants. Effects of type and concentration of sugar, light intensity and dark treatment on rooting and subsequent acclimatization of microshoots were studied. The significantly greater percentage (72-93%) of shoots formed roots on 0.5 mg L1 NAA, as compared to 55-73% on 0.5 mg L1 IBA. The maximum rooting (100%) and highest number of roots per shoot (4.7) were obtained at a Photon Flux Density (PFD) of 25 μmol m2 sec1 on the medium with 30 g L1 sucrose. Dark treatment during 7 days had significant positive effect on the root induction. For acclimatization in the greenhouse, rooted shoots were transplanted into different substrates containing peat, perlite and vermiculite at different ratios. Survival rate in 3 months was 96-100% and all the plants exhibited a normal phenotype. The developed technology may be used for mass propagation of common ash superior trees.