We administered estradiol and progesterone to spayed guinea pigs, with resultant accumulation of secretory granules in endometrial gland cells. By initially employing protein A-colloidal gold immunolocalization of relaxin, followed by cytochemical staining of carbohydrate with the thiocarbohydrazide-silver proteinate method on the same section, we showed clearly that the secretory granules were composed of a central core containing relaxin and a cortex of carbohydrate-rich material. Use of normal rabbit serum rather than relaxin antiserum, and omission of periodic acid, demonstrated the specificity of the technique.