Microscopic examination of cells in the sediment of voided urine is the principal non-invasive laboratory method of diagnosis, detection and follow-up of malignant epithelial tumors of the urothelium. Because of need for an objective interpretation of this difficult material, we initiated a program of computer-based image analysis of cells in the urinary sediment with the purpose of developing an automated diagnostic laboratory instrument. The potential practical significance of this approach was enhanced further because voided urine has proven unsuitable for analysis by flow cytometry. In preliminary studies, using visually selected cells, it was documented that image analysis could be reliably used to reach diagnostic conclusions, consistent with the cytologic diagnosis on the same material. During the current grant period we confirmed these results using unselected cells to reach computer-generated diagnostic results in 40 patients with a sensitivity of 95% and specificity of 88%. As the next step we developed a television based, semi-automated system of diagnostic cellimage analysis. A trial of 119 voided urine sediments from 103 sequential patients and 16 normal volunteers was conducted with most encouraging results. The performance of the system was comparable to routine cytology for high grade tumors and was superior for low grade tumors. The rate of """"""""false positive"""""""" results was high in patients with prostatic hypertrophy and was most likely due to co-existing urothelial abnormalities. We propose to test these results by image analysis of 600 cytospin samples in conjunction with cytology, histology and clinical findings. We also propose to accelerate image acquisition and analysis in order to create an instrument of practical laboratory value. The reasons why image analysis is capable of recognizing some low grade tumors not identified by human observers will be investigated on the premise that the cells of such tumors may have an abnormal DNA content or other features, recognized by the high resolution system. We propose to continue a study of DNA content and morphometric computer analysis of cells in urinary sediment to be compared with DNA ploidy of tumors and clinical data. DNA content of archival tumor material embedded in paraffin will be measured on nuclei obtained by a modification of Hedley's method. Because of known significance of aneuploidy of bladder tumors as a factor of grave prognostic significance, this aspect of our study will also permit the assessment of the prognostic value of image analysis of urinary sediments.
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