A cannulation and serial bleeding procedure has been developed to monitor the peripheral patterns of hormones associated with reproduction for up to 10 d in broiler breeder hens. Hens were cannulated via the jugular vein and returned to individual cages. The unrestrained cannulated hens were connected to a tether and swivel system that permitted constant infusion for maintenance of the cannula prior to serial bleeding and unrestrained long-term serial bleeding for up to 10 d. In a short-term experiment hens were bled every 12 min for 36 h, and in a long-term experiment hens were bled hourly for 10 d. In these experiments, 1.5-mL blood samples were collected at each time point with sodium citrate as the anticoagulant. To avoid hemodilution, after removal of plasma the red blood cells were reconstituted with saline to the original volume and returned to the hen of origin. Collection of serial blood samples was successful from 94% of hens in the short-term experiment and 79% of hens in the long-term experiment. Egg production was not affected (P > 0.05) during the 6 wk following serial bleeding in the short-term experiment. For hens that continued laying, egg production for 10 d prior to cannulation was not different (P > 0.05) from egg production for the 10 d during serial bleeding in the long-term experiment. However, late in the reproductive cycle many hens (25%) stopped laying when serially bled. It is concluded that this cannulation procedure can be used to study short-term or long-term peripheral patterns of hormones associated with oviposition and ovulation in laying broiler breeder hens.