In situ nucleic acid hybridization was applied to the detection of Chlamydia trachomatis on microscope slides by use of sulphonated total DNA as a probe. Visualization of labelled DNA was obtained using a commercial enzyme-linked monoclonal antibody. A mixture of paraformaldehyde and glutaraldehyde was found to be the best fixative. With high probe concentration (10 micrograms/ml), intracellular inclusions were detected as early as 8 h after inoculating the cell culture. Extracellular elementary bodies could also be detected. Five genital specimens were tested by in situ hybridization; the results were in agreement with those observed by culture.