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Objective: To establish the feasibility of a machine scoring method for her2/neu immunohistochemistry in samples of breast carcinoma., Study Design: A total of 65 consecutive cases of breast carcinoma with immunohistochemical stainingfor her2/neu by the Herceptest (Dako Corp., Carpinteria, California, U.S.A.) method (DAB chromogen with hematoxylin counterstain) were analyzed using an Extended Slide Wizard (Tripath Imaging, Inc., Burlington, North Carolina, U.S.A.) workstation running prototype software. Representative fields of view from the positive control, negative control and up to 10 fields from the stained tumor sample were captured interactively with a phased alternating line 3 CCD color camera. To determine the amount of specific membrane staining, chromogen separation of nuclear counterstain and membrane-positive stain was performed based on their respective absorption coefficients in the three color channels. The amount of specific membrane staining was scored based on a training set covering the rangefrom 0 to 3 + staining scores according to Dako. Manual scores of 2 + were tested for amplification by fluorescence in situ hybridization., Results: The automated scoring results correlated highly with the manual scores obtained per the Herceptest (Dako) instructions (R2>.92). The results were obtained in real time in the interactive mode., Conclusion: Machine scoring of immunohistochemical stains is practical, rapid and inherently reproducible, especially for samples with 1+ and 2+ manual scores.

Objective: To compare the diagnostic efficacy of a thin-layer technique with that of a membrane filter technique using a wide variety of fresh cytologic specimens., Study Design: Paired samples from 272 nongynecologic cytology specimens were processed for microscopy using thin-layer and membrane filter preparation techniques. Specimens included 162 body fluids and urines, 32 bronchial aspirates and 78 fine needle aspiration biopsies. The two techniques were compared for diagnostic efficacy, cell density/specimen, cell types recovered, qualitative cytomorphology, ease of interpretation and long-term stability of the final preparations., Results: Diagnoses supported by filter preparations were also supported by their matched thin-layer preparations. The same cell types were recovered by both techniques, and the slide cell density was slightly greater using membrane filters. Qualitative morphology and ease of use were superior with the thin-layer samples. After two years, all thin-layer slides were stable and readable, whereas only 43% of the membrane filter slides were usable., Conclusion: Thin-layer techniques are morphologically comparable and, in some ways, superior to membrane filter techniques for processing a wide variety of nongynecologic cytology specimens.

Objective: To determine the feasibility and utility of thin-layer cytology preparations for morphometric analysis of nuclear and cytoplasmic area in cells from nongynecologic cytology specimens., Study Design: Identical paired samples from nongynecologic specimens (bronchial aspirate, urine, pleural and peritoneal fluid) were used to prepare thin-layer preparations and corresponding traditional membrane filter preparations. The paired preparations were analyzed by static image morphometry measuring eight nuclear and cytoplasmic parameters that allowed calculation of nuclear area, cytoplasmic area and total cell size. Hepatocytes and bronchial columnar, mesothelial, squamous and transitional cells were studied, as was a single case of high grade transitional cell carcinoma. Sufficient numbers of each cell type were measured to allow statistical analysis., Results: Both thin-layer and membrane filter preparation techniques yielded individual cells suitable for morphometric analysis, and there were no consistent morphologic measurement differences between the two methods. The thin-layer preparation had the following significant technical advantages: more numerous easily measured single cells, lack of interfering background and superior specimen stability. Benign oval to round nuclei from a variety of cell types have a mean nuclear area in a narrow range from 29 to 55 microns 2. The mean nuclear area of malignant cells studied was significantly larger (78 microns 2), and there was a significant decrease in absolute cytoplasmic profile area in the malignant population studied., Conclusion: Thin-layer cytology preparations have significant advantages for morphometric studies over traditional membrane filter cytologic preparations. The morphometric measurement of nuclear area in benign and malignant cell populations has great potential as a generic screening tool for malignancy in cytologic specimens. Simultaneous measurement of cytoplasmic area adds a powerful dimension. The subsequent calculation of a true nuclear/cytoplasmic ratio may yield a sensitive and specific discriminator for detecting malignant cell populations in human nongynecologic cytology specimens.