His scientific work includes substantial contributions to the understanding of motion perception in insects and color vision in humans.
From 1939 to 1949, Hassenstein studied biology, physics, and chemistry in Berlin, Göttingen, and Heidelberg.
During his military service in 1943, he met Werner E. Reichardt, who eventually became his academic partner.
In 1948, he worked as an assistant at the Max Planck Institute for Marine Biology in Wilhelmshaven.
In 1958, Hassenstein worked with physicist Werner Reichardt and engineer Hans Wenking to found the world's first working group on cybernetics at the Max Planck Institute for Biology in Tübingen.