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As a preliminary step in the study of antibodies against spermatozoa in fertile and sterile women it was necessary to develop a highly sensitive technic capable of demonstrating the presence of small amounts of antibodies. ,4mong other serological tests the detection of anti-human sperm agglutinins offered a promising field, although previous investigators with mammalian spermatozoa had obtained only relatively low agglutination titers. Therefore, various methods of increasing the delicacy of the agglutination test by modilfying the standard technics were tried.These studies led to the development of a new type of best, the gelatin agglutinatilon teohnic, which is capable of detecting sperm agglutinins in titers about 100-fold higher than with the ordinary agglutination tests. The technic is described since this test may have possible application in the detection of agglutinins against other organisms than spermatozoa. The rationale of the test is based on the use #of live motile spermatozoa as antigens and the employment of a viscous medium in the form of 2.570 gelatin. This medium prevents the actively m'otile spermatozoa, once agglutinated, from disentangling themselves from the weakly bound floccules in high antiserum dilutions. I t also retards sedimentation, thus rendlering unnecessary redispersal of the delicately fliocculated sediment.Technic of test. Antigens. Donors with numerous motile spermatozoa and readily liquefying semen are used for the preparation od the antigen. Specimens of semen with large amounts of cellular debris are rejected to minimize nonspecific agglutination. -4 fresh ejaculum is allowed to stand until liquefaction is complete, usually in lfrom 30 to 60 minutes. A sperm count is done and the specimen is diluted to approximately 60 million sperma-Boston University School of J4edicin.e. tozoa per ml with physiological sodium chloride solution or Baker's buffered glucose solution( 1). The diluted semen is allowed to stand undisturbed for about an hour at room temperature to permit settling of clumped and immobile spermatozoa and cellular debris. The supernate, containing the actively motile spermatozoa, is carefully removed, a repeat sperm count is made, and the suspension is adjusted ttol 40 million spermatozoa per ml. An equal volume of gauze-filtered 10% gelatin in physiological sodium chloride soh tim is then added, giving a sperm concentration of 201 milIion per ml and a gelatin content of 570.The gelatin solution and gelatin-sperm mixture are maintained at aibout 36-31°C to facilitate pipetting.Test. Serial dilutions of the sera to be tested are made with physiological sodium chloride solution, the range depending upon the expected titers. A total of 0.3 ml of each dilution is first placed in a 65 x 5 rnm precipitation tube and an equal volume of the spermatozoa-gelatin antigen is then added giving a final concentratiion of 10 million spermatozoa per ml in 2.5% gelatin. The use of these small precipitation tubes not only conserves reagents, but also; provides a 30 mm column of flui...