[A] This publication was prepared by N. L. VANDEMARK, Professor of Dairy Physiology. Research reported herein was carried out cooperatively by the senior author and W. J. MILLER, W. C. KINNEY, JR., CARLOS RODRIGUEZ, and M. E. FRIEDMAN, formerly members of the Department of Dairy Science. The first announcement in 1951 of the successful impregnation of a cow with bull semen that had been frozen stimulated much interest and research in freezing as a method of preserving bull semen. Research during the years following 1951 resulted in considerable progress and success in using freezing as a means of holding semen for long periods of time without loss of fertility. Between 1952 and 1957, research on many aspects of the preservation of bull semen at sub-zero temperatures was carried out in the Department of Dairy Science at the University of Illinois. Many of these investigations have aided in perfecting the freezing technique that has been adapted for practical use. Some of these findings have been published, but many items have gone unreported except for general references at scientific meetings. It is the purpose of this bulletin to bring together the results of several experiments carried out in connection with the freezing of bull semen and to present a practical freezing procedure based on the results of these experiments and findings at other institutions. Persons interested in the development of the procedures and the reasons why certain steps are necessary in freezing semen will find the details in the first part of this bulletin. Those interested only in the freezing procedure may turn to page 35 where a practical method of freezing semen is described. EARLY WORK ON FREEZING SEMEN While it has been known for a long time that various types of tissues and organisms can withstand freezing and are even preserved by freezing, the first attempts at freezing sperm cells were made before the turn of the century. In 1897, Davenport[1][B] found that human sperm would withstand freezing. For thirty to forty years after that, little attention was paid to freezing as a possible means of preserving semen. An excellent review of the early attempts to freeze sperm has been assembled by Polge and Parkes.[2] These investigators also gave a good account of their work at the British National Institute of Medical Research in London, where in 1949 they demonstrated that glycerol would protect fowl sperm so that it would survive freezing. The next year they found that bull sperm and the sperm of several other species were protected by glycerol during freezing. During the same year, Emmens and Blackshaw[3] showed that ram and bull sperm would survive freezing. In 1951 frozen semen was used to produce a calf in England and a lamb in Australia. [B] These numbers refer to literature citations on page 38.