Bioincising is a biotechnological process to improve the permeability of wood by biological organisms. In recent years, there has been a great interest to determine the changes in the anatomical structure of wood as a result of bioincising process. In this study, the effects of bioincising by Physisporinus vitreus on the pit structure and permeability of Picea orientalis L. sapwood and heartwood were studied. Bioincised and non-bioincised samples were then treated with micronized copper quaternary and Celcure AC-500 wood preservatives by either dipping or vacuum method. The copper distribution and amount of copper retained in treated wood were evaluated by SEM-EDX and ICP-OES, respectively. The area of copper fixation in treated wood was also measured by the ArcGIS software package. The effects of P. vitreus activity were examined in the wood microstructure after bioincising by light microscope and SEM analyses on radial sections. After the bioincising, a significant increase was observed in the uptake of wood preservatives by vacuum, particularly in heartwood. The measurements on bordered pit, crossfield pit, and ray tracheid bordered pit dimensions in the microstructure of wood indicated that the degradation of pits was the most important factor in improved penetration and uptake of preservative solutions employed.