We measured the individual lengths of fluorescent labels on the three subdivisions of the endosteal envelope in iliac bone biopsy specimens produced by the administration of both oxytetracycline and demethylchlortetracycline. Fifty-one healthy subjects and 53 patients with postmenopausal osteoporosis were labeled in the stated order, and 8 osteopenic patients were labeled in the reverse order. Whatever the order of administration, the demethylchlortetracycline label was longer than the oxytetracycline label. We conclude (1) the difference in label lengths reflects a difference between the two compounds in some intrinsic property, whether physical, chemical, or pharmacokinetic. (2) If the calculation of extent of mineralizing surface is based on the mean length of the two labels, a suitable correction should be applied to the shorter label; alternatively, the length of the longer label alone should be used. (3) Unlabeled osteoid not due to label escape probably results from slow terminal mineralization after cessation of matrix synthesis during which too few tetracycline molecules are incorporated to exceed the threshold for visible fluorescence, rather than from the temporary interruption of mineralization followed by its resumption.