Objective: To investigate the hypothesis that image cytometry of sputum specimens can detect squamous carcinoma without requiring visually abnormal cells. • Design: The sensitivity and specificity of image cytometry were evaluated in a case-control study. • Material and Methods: Seventy-three sputum slides from the Mayo portion of the National Cancer Institute Cooperative Early Lung Cancer Study were restained by a modified Feulgen method. We examined 40 slides from 9 patients in whom squamous carcinoma developed and 33 slides from 11 patients in whom no cancer developed during a follow-up of at least 5 years. Images of normal epithelial nuclei were collected by using an automated image cytometer. Discriminant analysis was used to determine differences in DNA distribution of normal nuclei in sputum specimens from noncancer patients versus normal nuclei in sputum samples from patients in whom carcinoma developed. • Results: By using features based on DNA distribution, 74% correct classification of nuclei was possible without human review of the material and without the use of visually abnormal nuclei. A receiver operating characteristic curve demonstrated sensitivities and specificities, including 40% sensitivity and 90% specificity. • Conclusion: Although this study was limited to 20-year-old slides and squamous cell carcinoma, automated image cytometry detected a substantial proportion of patients with squamous cell cancer without using visually abnormal nuclei.
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