Fine-needle aspiration (FNA) has been used as a fast, minimally invasive, and reliable method for the evaluation of enlarged lymph nodes. However, there are some cases where the definitive diagnosis cannot be elicited with morphology alone, especially cases without a known primary lesion. Although immunocytochemical studies may be helpful in some situations, they are often complicated by nonspecific staining. Recently, a novel tyramide-based tyrosinase assay was developed. Since melanocytes, both benign and malignant, produce tyrosinase, we postulated that this assay could be useful as an in situ biochemical diagnostic test. We modified the Perkin Elmer TSA assay, a commercial assay based on tyramide, a tyrosine analog that is a substrate for tyrosinase, for use on air-dried cytological preparations. We validated the assay on cell lines, then tested a small series of melanoma and nonmelanoma cytology specimens. The YUGEN8 melanoma cell line was used to optimize the assay and it showed abundant reaction product, while HeLa cells served as a negative control. All melanoma cytology specimens were positive and all nonmelanoma specimens were negative. These results suggest that this simple, fast, and inexpensive assay is a sensitive and specific method for detection of melanoma cells in cytology specimens. This method may be a useful ancillary procedure for the resolution of challenging melanoma cases.
Despite low sensitivity (around 60%), cytomorphologic examination of urine specimens represents the standard procedure in the diagnosis and follow-up of bladder cancer. Although color is information-rich, morphologic diagnoses are rendered almost exclusively on the basis of spatial information. We hypothesized that quantitative assessment of color (more precisely, of spectral properties) using liquid crystal-based spectral fractionation, combined with genetic algorithm-based spatial analysis, can improve the accuracy of traditional cytologic examination. Images of various cytological specimens were collected every 10 nm from 400 to 700 nm to create an image stack. The resulting data sets were analyzed using the Los Alamos-developed GENetic Imagery Exploitation (GENIE) package, a hybrid genetic algorithm that segments (classifies) images using automatically 'learned' spatio-spectral features. In an evolutionary fashion, GENIE generates a series of algorithms or 'chromosomes', keeping the one with best fitness with respect to a user-defined training set. First, we tested the system to determine if it could recognize malignant cells using artificial cytology specimens constructed to completely avoid the requirement for human interpretation. GENIE was able to differentiate malignant from benign cells and to estimate their relative proportions in controlled mixtures. We then tested the system on routine cytology specimens. When targeted to detect malignant urothelial cells in cytology specimens, GENIE showed a combined sensitivity and specificity of 85 and 95%, in samples drawn from two separate institutions over a span of 4 years. When trained on cases initially diagnosed as 'atypical' but with unequivocal follow-up by biopsy, surgical specimen or cytology, GENIE showed efficiency superior to the cytopathologist with respect to predicting the follow-up result in a cohort of 85 cases. We believe that, in future, this type of methodology could be used as an ancillary test in cytopathology, in a manner analogous to immunostaining, in those situations when a definitive diagnosis cannot be rendered based solely on the morphology. Laboratory Investigation ( Keywords: atypical; urine; bladder; cytopathology; genetic algorithm Urothelial carcinoma is diagnosed and followed by cytological evaluation of voided urine and bladder washings.1,2 The sensitivity of these tests is moderate, ranging between 60 and 80%.3,4 Low-grade tumors offer an even greater challenge to the cytopathologist, 5 resulting in the use of such terminology as 'atypical urothelial cells', leading to suboptimal patient management. 6 Morphologic diagnoses in cytopathology are primarily driven by spatial relationships, while colorbased information is relegated to a very minor role. This may be because the human eye has limited spectral resolution, viewing the world in broad, overlapping spectral bands of red, green and blue. If the spectral properties of cytological specimens are important, this fact can only be assessed using devices capable of imaging with...
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