The procedure of wood sample preparation, including tree-ring dissection, cellulose extraction, homogenization and finally weighing and packing for stable isotope analysis is labour intensive and time consuming. We present an elaborated methodical guideline from pre-analyses considerations, wood sample preparation through semi-automated chemical extraction of cellulose di- rectly from tree-ring cross-sections to tree-ring dissection for high-precision isotope ra- tio mass spectrometry. This guideline reduces time and maximizes the tree-ring stable isotope data throughput significantly. The method was applied to ten different tree species (coniferous and angiosperm 10 wood) with different wood growth rates and differently shaped tree-ring boundaries. The tree-ring structures of the cellulose cross-sections largely remained well identifiable. FTIR (Fourier transform infrared) spectrometry and the comparison of stable isotope values with classical method confirm chemical purity of the resultant cellulose. Sample homogenization is no longer necessary. Cellulose extraction is now faster, cheaper and more user friendly allowing (i) the simultaneous treatment of wood cross-sections of a total length of 180 cm (equivalent to 6 increment cores of 30 cm length) and thickness of 0.5 to 2 mm, and (ii) precise tree- ring separation at annual to high-resolution scale utilizing manual devices or UV-laser microdissection microscopes.