This protocol describes a method for the in vivo measurement of steroid hormones in brain circuits of the zebra finch. A guide cannula is surgically implanted into the skull, microdilysate is collected through a microdialysis probe that is inserted into the cannula, and steroid concentrations in the microdialysate are determined using the enzyme-linked immunosorbent assay (ELISA). In some cases, the steroids measured are derived locally (e.g., neural estrogens in males), whereas in other cases, the steroids measured reflect systemic circulating levels and/or central conversion (e.g., the primary androgen testosterone and the primary glucocorticoid corticosterone). A reverse-microdialysis (retrodialysis) method that can be used to deliver pharmacological agents into the brain to influence local steroid neurochemistry as well as behavior is also discussed.