This book constitutes the refereed proceedings of the 16th Scandinavian Conference on Image Analysis, SCIA 2009, held in Oslo, Norway, in June 2009. The 30 revised full papers and 49 revised poster papers presented were carefully reviewed and selected from 154 submissions. The papers are organized in topical sections on human motion and action analysis, object and pattern recognition, color imaging and quality, multispectral color science, medical and biomedical applications, image and pattern analysis in astrophysics and astronomy, face recognition and tracking and computer vision.