Lung cancer remains the leading cause of cancer deaths in the developed world. There is no widely accepted method to screen for this cancer. The most commonly used method remains conventional sputum cytology, but this method is hampered by low sensitivity. We tested the hypothesis that sensitivity of sputum cytology for early lung cancer can be greatly improved by using image analysis of sputum cells, at a modest reduction of specificity.The study was double-blinded and used sputum samples from subjects with well-characterized clinical diagnoses. There were 177 cancers, 98 dysplasias, and 558 normals. The study samples were separated into two independent sets: training set and test set. Sputum samples were collected prospectively from subjects with a high probability of having lung cancer. Seven institutions from five countries participated in the study. All subjects had complete clinical diagnoses which included, as a minimum, negative chest x-rays for all negative cancers, while all cancers had confirmed tissue pathology. Samples were prepared according to the Saccomanno method. For conventional cytology, slides were stained using Papanicolaou stain. For image analysis, slides were stained using a DNA-specific (Feulgen-Thionin) stain. An automated, high-resolution image cytometer was used for measurements.At 90% specificity, sensitivity of 60% can be achieved for adenocarcinoma, compared to only 14% sensitivity of conventional cytology (at 99% specificity). Similarly, 45% sensitivity at 90% specificity can be reached for stages 0 and I lung cancer, compared to only 14% (at 99% specificity) using conventional cytology.Cytometry combined with conventional cytology shows an increase in sensitivity to early-stage cancer and to adenocarcinomas compared to conventional cytology alone. While the results are encouraging, the sensitivity to detect early lung cancer should be further improved to 70 -80% at 90 -95% specificity before this test can be considered for screening of high-risk individuals for lung cancer. Cytometry (Clin. Key terms: lung cancer; sputum cytology; image analysis; sensitivity; specificity; screening for lung cancer; early lung cancer; adenocarcinoma Despite the fact that advances have been made in sputum cytology over the last 30 years (1,2), the literature contains mixed results about the efficacy of using it as the primary method for detection of lung cancer. Although approximately 50 studies employing a range of preparation techniques reported sensitivities ranging from 40 -80% at high specificities (95-99%) (3), they suffer from a limited number of samples and are demonstrated by latestage cancer. Large-scale studies, such as the NCI-sponsored study (the Cooperative Early Lung Cancer Detection Program, carried out by Memorial Sloan-Kettering, the Mayo Clinic, and Johns Hopkins University) (4 -6), did not confirm such results despite the fact that the most experienced cytopathologists of the day were carrying out the sputum cytology. Sputum cytology alone detected only 23% of the cancers in th...