The goal of this study was to determine if selected biochemical, structural and spectral properties of Norway spruce needles are influenced by the azimuth orientation of the branch. Three youngest needle age classes from 20 mature (100 years old or older) Norway spruce trees were sampled from upper branches of the sunlit production crown part from each of the 4 cardinal azimuth orientations. Photosynthetic pigments, soluble phenolic compounds and selected spectral and structural characteristics were determined for each needle age class. The content of photosynthetic pigments and soluble phenolic compounds did not differ among needles from different azimuth-oriented branches, nor did the optical reflectance indices Normalized Difference Vegetation Index (NDVI), Transformed Chlorophyll Absorption in Reflectance Index (TCARI)/Optimized Soil-Adjusted Vegetation Index (OSAVI), Red Edge Inflection Point (REIP) and Landsat Thematic Mapper bands 5 and 4 (TM5/TM4). No variation in volume properties, tissue volume proportions and cross-section shape characteristics of 3rd-year needles rejected our hypothesis that there would be variation in needle structural properties according to the azimuth orientation of branches. Consequently, we concluded that a random sampling of similar-aged needles within the sunlit production crown part may be used to study biochemical or structural and spectral needle properties of a mature Norway spruce growing in forest stands without a significant slope. In addition, the results obtained from a branch of one azimuth orientation should be representative for the whole sunlit portion of the crown. Consequences of these findings for Norway spruce health monitoring using remote sensing techniques are discussed.