Karl von Frisch was the son of the surgeon and urologist Anton von Frisch (1849-1917), by his marriage to Marie Exner. Karl was the youngest of four sons, all of whom became university professors.

Karl studied in Vienna under Hans Leo Przibram and in Munich under Richard von Hertwig, initially in the field of Medicine, but later turned to the natural sciences. He received his doctorate in 1910 and in the same year started work as an assistant in the zoology department of the University of Munich.

In 1912 he became a lecturer in zoology and comparative anatomy there; and in 1919 was promoted to a professorship. His research on honeybees was continued by his student Ingeborg Beling. In 1921 he went to Rostock University as a professor of zoology and Director of an institute. In 1923 he accepted the offer of a chair at Breslau University, returning in 1925 to Munich University, where he became the head of the institute of zoology.

His work centered on investigations of the sensory perceptions of the honey bee and he was one of the first to translate the meaning of the waggle dance. His theory, described in his 1927 book Aus dem Leben der Bienen (translated into English as The Dancing Bees), was disputed by other Scientists and greeted with skepticism at the time. Only much later was it shown to be an accurate theoretical analysis.

In 1933 the Nazi regime passed the Civil Service Law, requiring all public servants to provide proof of Aryan ancestry. Frisch was unable to account for the ancestry of one of his grandparents, and was therefore Classified as a mischling of 1/8th Jewish ancestry, but formally allowed to keep his job. However groups of students and lecturers worked to have him dismissed from the university, preferring a committed National Socialist. Frisch also attracted negative attention for employing Jewish assistants, including many women, and for practicing "Jewish science". Eventually Frisch was forced into retirement, but the decision was reversed due to advances in his research on combating nosema infections in bees and his forced retirement was postponed until after the war. Frisch also worked actively to help Polish Scientists who had been singled out for internment by the Gestapo.

The institute of zoology was destroyed in the Second World War, and in 1946 Frisch went to work at the University of Graz, remaining there until 1950, when he returned to the Munich institute after it was reopened. He retired in 1958 but continued his research.

Karl von Frisch married Margarete, née Mohr, who died in 1964. Their son, Otto von Frisch, was Director of the Brunswick natural history museum between 1977 and 1995.

The linguistic findings described above were based on Frisch’s work primarily with the Carnica variety of bees. Investigations with other varieties led to the discovery that language elements were variety-specific, so that how distance and direction information is relayed varies greatly.