The manipulation of single molecules is still in its infancy but significant breakthroughs have been made in chemistry, biology, physics, engineering, and polymer science. Demonstrating the potential of atomic force microscopy (AFM) beyond imaging, this book discusses concepts and methodologies used to manipulate and study single (bio)molecules with AFM. The first part of the text is dedicated to the pulling of single molecules with force spectroscopy to investigate molecular interactions, mechanics, and mechanochemical processes. The second part addresses the manipulation, repositioning, and targeted delivery of single molecules on substrates.