The effects on morphology and diagnostic interpretation of delayed processing of refrigerated effusion samples have not been well documented. The potential for cellular degeneration has led many laboratories to reflexively fix samples rather than submit fresh/refrigerated samples for cytologic examination. We sought to determine if effusion specimens are suitable for morphologic, immunocytochemical, and DNA-based molecular studies after prolonged periods of refrigerated storage time. Ten fresh effusion specimens were refrigerated at 4 degrees C; aliquots were processed at specific points in time (days 0, 3, 5, 7, 10, 14). Specimens evaluated included four pleural (3 benign, 1 breast adenocarcinoma) and six peritoneal (2 ovarian adenocarcinomas, 1 malignant melanoma, 2 mesotheliomas, 1 atypical mesothelial) effusions. The morphology of the cytologic preparations from the 10 effusions was preserved and interpretable after 14 days of storage at 4 degrees C. The immunocytochemical profile of the samples (AE1/AE3, EMA, calretinin, and LCA) was consistent from day 0 to day 14. Amplifiable DNA was present in all samples tested on day 14. We conclude that cytopathologic interpretation of effusion samples remains reliable with refrigeration at 4 degrees C even if processing is delayed.
BACKGROUND Squamous atypia in postmenopausal (PM) cervical vaginal smears (CVS) only rarely is associated with a biopsy‐proven squamous intraepithelial lesion (SIL), and thus most commonly represents an atrophy‐associated benign reactive change. METHODS To distinguish atypical squamous cells of undetermined significance (ASCUS) and SIL from atrophy‐associated benign reactive changes, a review of atypical atrophic PM CVS was performed. Ninety CVS exhibiting an atrophic smear pattern were considered appropriate for study. Repeat smears and/or biopsy after local estrogen therapy were requested to distinguish atrophic/reactive from dysplastic changes. RESULTS Generally, atrophic CVS exhibit uniform nuclear enlargement in the squamous cell population, which, using the criterion of nuclear enlargement alone, would qualify the majority of these cases to be classified as ASCUS. The nuclear enlargement associated with atrophy resolves with the local application of estrogen. Follow‐up after local estrogen treatment was available for 84 of 90 patients and revealed 10 cases of SIL (12%) and 9 cases of ASCUS (11%), 6 of which were favored to be of a reactive etiology. Nuclear features most commonly noted in the cases considered to be ASCUS (nonreactive) and SIL were nuclear hyperchromasia and nuclear contour irregularities. CONCLUSIONS Nuclear enlargement alone is not sufficient for diagnosing ASCUS or SIL in PM CVS. Nuclear enlargement in squamous cells is an expected normal reactive change present in PM CVS that resolves with the application of local estrogen. Nuclear hyperchromasia and irregular nuclear contours remain the most reliable cellular characteristics for diagnosing SIL in atrophic CVS. [See editorial on pages 200‐1, this issue.] Cancer (Cancer Cytopathol) 1998;84:218‐225. © 1998 American Cancer Society.
the most accurate diagnostic results. The MART-1 antigen, a transmembrane protein , is specifically expressed in melanocytes and MM. A recently developed mono-effusion samples with antibodies to MART 1, HMB45, S-100, and cytokeratins (AE1/ AE3).IM stains were performed on cell block or cytospin material, depending on availability. In the morphologic review, emphasis was placed on Diff Quik-stained material, due to its enhanced cytoplasmic volume and detail. RESULTS. Predominant cytologic features noted were lack of cellular cohesion (in 100% of cases), large eccentric nuclei with prominent nucleoli (in 100%), multinu-cleation (in 84%), variable cytoplasmic vacuolization (in 75%), pigment (in 72%), and cell-in-cell engulfment (in 47%). All immunoreactive cases with sufficient material stained with at least one of the markers used. Tumor cells were positive with IM stains to MART-1 in 78% of cases, HMB45 in 81%, and S-100 in 81%. Coexpres-sion of MART-1, HMB45, and S-100 was noted in 63% of cases. Of cases that showed expression for only 1 of the 3 antigens, the MART-1 was positive in 1 case, and HMB45 and S-100 were positive in 2 cases each. Three cases showed immunoreactivity for cytokeratins in the melanoma cells. CONCLUSIONS. The diagnosis of MM in effusions can be made reliably through a combination of morphologic and IM features. Differential diagnosis often involves distinguishing MM from adenocarcinoma or reactive mesothelial cells. Cytoplasmic vacuo-lization, multinucleation, prominent nucleoli, and cell-in-cell engulfment are cytologic features common to all three. The lack of IM staining for cytokeratins alone cannot reliably distinguish MM; 11% of cases showed positive staining with this antibody in the melanoma cells. The use of a panel of antibodies increases the accuracy of diagnosing MM. In this study, MART-1 proved a useful adjunct to the HMB45/S-100/cytokeratin panel for the diagnosis of MM in effusions, staining 78% of the immunoreactive cases, with positivity in 1 case that was negative for HMB45 and S-alignant effusions associated with metastatic melanoma (MM) may exhibit a variable cytologic appearance. 1-5 The diagnosis
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