Honeybee (Apis meillfera) semen was stored for 359 days at -196°C in liquid nitrogen. Eleven semen diluents were tested. Use of a mixture of 40% semen and 60% Kiev diluent containing 10% dimethylsulphoxide, combined with freezing of samples at the rate of 3-4°/min produced relatively good post-thaw sperm motility. Fifteen queens were inseminated instrumentally with semen stored for 305-359 days. Thirteen survived and produced a brood area of 345 ± 60 cm2, of which 47'3 ± 6-42% were worker progeny from the frozen spermatozoa. Four queens were superseded by their daughters which mated naturally and built up strong colonies.