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PubMed1995

DNA index and ploidy distinguish normal human parathyroids from parathyroid adenomas and primary hyperplastic parathyroids.

BACKGROUND: The goal of this study was to identify factors that might aid in diagnosis and intraoperative management of hyperparathyroidism. METHODS: We analyzed biopsy specimens of 242 parathyroids from 159 patients by use of flow cytometry and image cytometry (ICM) for DNA index (DI), defined as the content of nuclear DNA compared with that expected for a DNA diploid standard, for proliferative index (PI), and for ploidy (diploid versus aneuploid or tetraploid). RESULTS: True normal and normal parathyroids from patients with solitary adenomas were uniformly diploid. Abnormal ploidy (aneuploidy or tetraploidy) was identified frequently in adenomas and occasionally in hyperplasias with the exception that multiple endocrine neoplasia (MEN) biopsy specimens were uniformly diploid. DI for adenomas was similar to that for hyperplasias, and DI of both was higher than for normal glands. ICM-DI correlated positively with flow cytometry-DI and patient age and inversely with serum parathyroid hormone. PI was relatively low in all groups but was higher for hyperplasias versus normal parathyroids from patients with solitary adenomas and MEN versus non-MEN. PI correlated inversely with patient age. CONCLUSIONS: DI by ICM differentiates normal from abnormal parathyroids. DI might influence extent of resection in two- and three-gland hyperplasia and selection of the most appropriate gland for autografting and cryopreservation in patients with four-gland hyperplasia.

Adenoma↗
PubMed1993

Molecular and biological factors in the pathogenesis of ovarian cancer.

BACKGROUND: The classic prognostic parameters are insufficient for predicting the prognosis of the individual patient. Knowledge of molecular and biological factors which are responsible for the development and progression of ovarian cancer may improve the prediction of prognosis. MATERIALS AND METHODS: Recent data both on factors associated with the development and control of ovarian cancer cells and on DNA ploidy have been reviewed. RESULTS: Observations suggest that steroid and peptide hormones have a role in disease etiology and progression, and that peptide growth factors and cytokines, oncogenes and tumor suppressor genes, by their impact on mitosis and cell number may influence the rate of mutations, which could confer malignant transformation. DNA ploidy is an objective independent prognostic factor. DNA aneuploidy indicates high risk, diploidy low risk. Only tumours shown to be DNA diploid by flow-cytometry and image cytometry are considered diploid. S-phase fraction is currently not reliable. CONCLUSION: Understanding the mechanisms involved in ovarian cancer development and growth will allow opportunities for the rational design of effective anti-tumour treatment modalities. More objective and reproducible prognostic variables will improve the predictiveness of prognosis.

DNA, Neoplasm↗
PubMedDate not supplied

Signet ring cell carcinoma of the stomach: a morphometric, ultrastructural, and DNA cytometric study.

The present study was undertaken to determine whether or not signet ring cell (diffuse, isolated cell) gastric carcinomas display a specific profile at the ultrastructural, morphometric, and DNA cytometric levels. Thirty-two cases of gastric carcinoma and 8 cases of peptic ulcer (control group) were studied with electron microscopy, morphometry, flow cytometry, and image cytometry. Despite the ultrastructural cellular heterogeneity of signet ring cell carcinomas, the neoplastic cells display fairly constant morphometric features: The cellular and nuclear volumes are significantly smaller than those of the other types of gastric carcinomas and closely resemble those of normal foveolar cells. The relatively small size of signet ring cell carcinoma nuclei fits with the high percentage of the cases of this type of gastric carcinoma that are either diploid or nearly diploid. There is a relationship between the infiltrative pattern of growth of gastric carcinoma (regardless of histologic subtype and ultrastructural cell differentiation) and the small size of neoplastic cells and their nuclei.

Adenocarcinoma, Mucinous↗
PubMed2001

Cross-presentation by dendritic cells of tumor antigen expressed in apoptotic recombinant canarypox virus-infected dendritic cells.

We have investigated the possible usefulness of recombinant canarypox virus (ALVAC) encoding the melanoma-associated Ag, Melan-A/MART-1 (MART-1), in cancer immunotherapy, using a dendritic cell (DC)-based approach. ALVAC MART-1-infected DC express, and are able to process and present, the Ag coded by the viral vector. One consistent feature of infection by ALVAC is that these viruses induce apoptosis, and we show cross-presentation of Ag when uninfected DC are cocultured with ALVAC MART-1-infected DC. Uptake of apoptotic virally infected DC by uninfected DC and subsequent expression of tumor Ag in the latter were verified by flow cytometry analysis, image cytometry, and confocal microscopy. Functional activity was monitored in vitro by the stimulation of a MART-1-specific cytotoxic T cell clone. Heightened efficiency in Ag presentation is evidenced in the 2- to 3-fold increase in IFN-gamma production by the T cell clone, as compared with the ALVAC-infected DC alone. Cocultures of ALVAC MART-1-infected and uninfected DC are able to induce MART-1-specific T cell immune responses, as assessed by HLA class I/peptide tetramer binding, IFN-gamma ELISPOT assays, and cytotoxicity tests. Overall, our data indicate that DC infected with recombinant canarypox viruses may represent an efficient presentation platform for tumor Ags, which can be exploited in clinical studies.

Antigen Presentation↗
PubMed1991

Automated analysis of rat breast tumors. Comparison of four methods.

Chemically-induced malignant rat breast tumors pose diagnostic dilemmas since the majority are well-differentiated, noninvasive papillary lesions that are barely distinguishable from benign papillary lesions. This study compared several automated modalities to see which best separated benign from malignant breast tumors. Thirty-three carcinogen-induced rat breast tumors (13 adenomas, 10 papillary carcinomas and 10 invasive carcinomas) were evaluated by static (image) cytometry (ICM) of integrated optical density, by flow cytometry (FCM) and by two automated morphometric protocols, contextual analysis and single-gland analysis. DNA ploidy analysis, by either ICM or FCM, did not discriminate between the benign and malignant tumors. Contextual analysis correctly identified 11 of 13 benign and 17 of 20 malignant lesions (P less than .01). Single-gland analysis correctly identified all 13 benign and 17 of 20 malignant lesions (P less than .01). No method distinguished invasive from noninvasive carcinomas. The data suggest that architectural features are more important than nuclear features in differentiating benign from malignant rat breast tumors.

Animals↗
PubMed1994

Seminomas of the canine testis. Counterpart of spermatocytic seminoma of men?

BACKGROUND: Dogs develop germ cell tumors of the testis at a relatively high rate. It is not known to what degree these tumors resemble various human testicular neoplasms. EXPERIMENTAL DESIGN: The epidemiology and morphology of a series of spontaneous canine testicular tumors, collected between 1985 and 1991, was analyzed, and compared with human testicular germ cell tumors. DNA content analysis of representative samples was performed using flow cytometry and image cytometry. Eight human spermatocytic seminomas were studied in parallel. RESULTS: All canine tumors had the histopathologic features reported as typical for dog testis seminomas. These tumors could show both an intratubular and an invasive component. Most of them were pure (78%), while they could be combined with a Leydig cell tumor, a Sertoli cell tumor, or both. No somatic, placental or yolk sac cells were identified, and there was no carcinoma in situ (CIS). A bimodal age distribution, with a peak around 1 year of age and between 4 and 16 years of age, was found for all pure and mixed testicular tumors, except for those composed of a Leydig cell and a seminoma component. These tumors were all present in dogs older than 7 years, being significantly more older (p < 0.01) than dogs with a pure tumor of either type. All Sertoli cell and Leydig cell tumors were diploid. No consistent peritriploid DNA content, characteristic of human testicular germ cell tumors, was found for canine seminomas, which most often had a diploid DNA content. Human spermatocytic seminomas always contained diploid tumor cells, and showed a relatively low number of high ploidy cells, comparable to canine seminomas of the testis. CONCLUSIONS: The so-called seminomas of the testis are tumors of old age. Histologically, these tumors are composed of a single cell type with some variation without evidence of differentiation. It is proposed that canine seminoma correspond to human spermatocytic seminomas. It is thought that the Leydig elements in these tumors represent a reactive change rather than biphasic differentiation of a single stem cell capable of germinal and sex-cord cell development.

Animals↗
PubMed2000

Evaluation of DNA ploidy and degree of DNA abnormality in benign and malignant melanocytic lesions of the skin using video imaging.

BACKGROUND: Making a morphologic distinction between benign and malignant melanocytic tumors of the skin is frequently difficult, especially because "gray zones" between these lesions often exist. DNA image cytometry as an adjuvant method for the diagnosis and prognostic prediction of premalignant lesions and malignant tumors of many other organs is already well established. The aim of this study was to determine whether DNA image cytometry is helpful in distinguishing benign from malignant melanocytic lesions and whether cytometry would give valid information with which to predict the prognoses associated with malignant melanomas. METHODS: DNA image cytometry was performed on 127 benign and 58 primary maligant melanomas of the skin as well as 11 metastatic melanomas, using an enzymatic single cell solution according to a method described by Heiden et al. in Cytometry (1991;12:614-21). RESULTS: DNA aneuploidy was graded by DNA index (DI) and a 2c deviation index (2cDI). In contrast to benign melanocytic lesions (with 16% DNA aneuploidy), primary and metastatic malignant melanomas had significantly higher frequencies of DNA aneuploidy (86% and 73%, respectively). In the degree of DNA aneuploidy, significant differences between benign and malignant melanocytic tumors could be observed. The mean 2cDI of aneuploid benign lesions was 1.0, whereas the primary malignant melanomas had a mean 2cDI of 2.92 and the metastatic melanomas a mean of 6.9. The frequency of DNA aneuploidy increased with Breslow thickness. Twenty-one patients with primary malignant melanoma developed metastases. All metastasizing primary tumors were aneuploid and showed a significantly higher grade of DNA aneuploidy than nonmetastasizing malignant melanomas. Moreover, none of the diploid malignant melanomas developed metastases. CONCLUSIONS: This study reveals that DNA image cytometry is prognostically and diagnostically relevant to the evaluation of melanocytic lesions of the skin. Nevertheless, it cannot be relied on alone to provide enough information for a diagnosis.

Adult↗
PubMed2003

Comparison of flow cytometry and image morphometry in the quantitative analysis of cell population markers in the lymph node of sheep.

Two approaches to the quantitative analysis of cell population markers in tissues are flow cytometry and image morphometry. To compare these methods, sheep lymph nodes were collected and analysed for CD8+ and CD21+ cell populations, which were selected to represent dispersed and concentrated cell populations, respectively. These two populations were measured as a percentage of total cell count (flow) or total tissue area (morphometry). The two populations were also measured as a percentage of respective base populations (CD2+ cells for CD8 and MHC II+ cells for CD21). A simple linear regression analysis showed that when the cell populations were assessed as a percentage of total cell count or total area, measurements obtained with flow and morphometry only correlated significantly with the dispersed CD8+ population and not with the highly concentrated CD21+ population. However, when the cell populations were assessed as a percentage of their base population, measurements obtained with flow and morphometry showed a significant correlation for both the dispersed and concentrated cell populations. This study demonstrates that measurements of lymph node cell populations obtained with the two methods are comparable, but that tissue distribution of cell populations should be considered, when the unit of measurement is chosen.

Animals↗
PubMed2004

[Noninvasive brush biopsy as an innovative tool for early detection of oral carcinomas].

PURPOSE: The aim of this prospective study was to investigate the diagnostic accuracy of DNA image cytometry in combination with non-invasive brush biopsies taken from suspicious oral lesions. MATERIAL AND METHODS: Cytological diagnoses obtained from 1328 exfoliative smears of 332 different lesions were compared with histology and/or clinical follow-ups of the respective patients. Additionally, nuclear DNA contents were measured after Feulgen restaining using a TV image analysis system. DNA aneuploidy was assumed if abnormal DNA stemlines or cells with DNA content greater than 9c were observed. RESULTS; The sensitivity of our cytological diagnosis in addition to DNA image cytometry on oral smears for the detection of cancer cells was 97.8%, specificity 100%, positive predictive value 100%, and negative predictive value 98.1%. CONCLUSION: The application of DNA image cytometry with DNA aneuploidy as a marker for neoplastic transformation in oral smears secures cytologic diagnosis of carcinomas. Smears from brushings of all visible oral lesions are an easily practicable, cheap, noninvasive, painless, and safe screening method for detection of oral precancerous lesions and squamous cell carcinoma in all stages. We conclude that DNA image cytometry is a very sensitive and highly specific, objective, and reproducible adjuvant tool for identification of neoplastic cells in oral smears.

Aged↗
PubMed1999

Cytologic and DNA cytometric diagnosis and therapy monitoring of squamous cell carcinoma in situ and malignant melanoma of the cornea and conjunctiva.

OBJECTIVE: To investigate the diagnostic accuracy of exfoliative cytology of the cornea and conjunctiva and DNA image cytometry for quality control and monitoring of therapy for malignant neoplasms. STUDY DESIGN: Conjunctival or corneal smears from six cases clinically suspicious for malignant melanomas and eight suspicious for carcinomas in situ were investigated. Smears from 18 cases clinically nonsuspicious for neoplastic diseases served as negative controls. Repeated smears were obtained during and after local mitomycin C (MMC) therapy. RESULTS: In none of 18 nonsuspicious cases, cytology revealed abnormal cells. DNA cytometry showed nonaneuploidy in all of these. All smears from patients with histologically proven malignant melanomas (MM) and squamous cell carcinomas in situ revealed abnormal cells. Image cytometry demonstrated DNA aneuploidy in 66.6% of patients with MM and 80% with carcinoma. Sensitivity of cytology thus was 100% for both MM and carcinoma; specificity also was 100%. DNA measurements after MMC therapy revealed euploid polyploidization of nonneoplastic squamous cells. DNA cytometry provided an objective identification of tumor cell regression. CONCLUSION: Cytologic examination of corneal and conjunctival smears is a noninvasive tool with high diagnostic accuracy for detection of epithelial neoplasms. DNA image cytometry can serve for quality control and for objective monitoring of the effect of local chemotherapy.

Aneuploidy↗
PubMed2001

Prognostic significance of DNA cytometry in carcinoma of the uterine cervix FIGO stage IB and II.

OBJECTIVE: To assess the prognostic value of DNA-image cytometry in cervical carcinoma of the uterus and its relation to other established prognostic factors. STUDY DESIGN: The study included 116 cases of cervical carcinoma FIGO stages IB and II which were treated with radical abdominal hysterectomy. The median follow-up was 55 months (range 1-162 months). DNA image cytometry was performed on cytologic specimens prepared by enzymatic cell separation from formalin-fixed, paraffin-embedded tissues. DNA stemline ploidy, DNA stemline aneuploidy, 5c exceeding rate, 9c exceeding rate, 2c deviation index, and DNA malignancy grade were computed. DNA-variables as well as various clinical and histological variables were related to survival rates. RESULTS: In multivariate statistical analysis DNA stemline ploidy using 2.2c as a cut-off value and FIGO stage showed to be statistically significant available presurgery predictors of survival, whereas the postsurgical parameters lymphonodal status, tumor size and parametrial involvement were significantly correlated with survival. The synopsis of all parameters in a multivariate Cox model indicated that - with declining relevance - the number of positive pelvic lymph nodes, DNA stemline ploidy using a cut-off level at a modal value of 2.2c, largest pelvic lymph node, 5c exceeding rate, and ratio of carcinoma area to cervix area, were of predictive value for survival. CONCLUSIONS: Our results suggest that prognostic information deducted from classical staging parameters is successfully complemented by DNA image cytometry which can be applied pretherapeutically.

Adult↗
PubMed1994

Comparative DNA analysis of breast cancer by flow cytometry and image analysis.

Measurement of DNA ploidy can be performed either with Flow Cytometry (FCM) and Image-Analysis (IA); both methods provide prognostic information in primary breast cancer. We compared the results of quantitative DNA analysis of formalin fixed, paraffin embedded tissue from 62 invasive ductal breast cancers. For FCM nuclear suspensions from disaggregated tumor were stained with Propidium Iodide and analyzed by means of Ortho Cytoron Absolute. For IA nuclear suspensions were stained by the Feulgen method and analyzed by means of Vidas system. We found a good correlation between flow cytometry DNA Index and Histogram Type, according to Auer classification (rs = 0.65, p < 0.001) and between DNA Index and Grading of Malignancy (MG) which had been measured by Bocking's algorithm (rs = 0.38, p < 0.05). Concerning to disease free survival (DFS), flow cytometric DNA Index showed a better correlation (rs = 0.56, p < 0.001). We concluded that the two methods provide comparable results, but offer individual advantages and are complementary for analyzing DNA ploidy in breast cancer.

Algorithms↗
PubMed1996

Application of image analysis cytometry in follicular fluid cells obtained from in-vitro fertilization cycles: relationships to patient's age, oocyte maturity, fertilizability and in-vitro fertilization outcome.

In an in-vitro fertilization (IVF)/embryo transfer programme granulosa cells obtained from 59 individual preovulatory follicles were analysed using multiparameter image analysis cytometry, in an attempt to determine whether their morphometric and DNA-cytometric parameters could prove useful in assessing follicle and oocyte maturity and in predicting fertilizability and outcome of these IVF cycles. Almost all morphometric and DNA-cytometric parameters were not correlated with either the patient's age or oocyte maturity, and did not predict oocyte fertilization or occurrence of a clinical pregnancy. The only possible relevant parameter which, despite its inverse correlation to total luteinizing hormone administration, also proved to be inversely correlated to pregnancy outcome (in the seven cases in which a pregnancy occurred), was the percentage of granulosa cell nuclei with increased DNA content (> 5c). Finally, if granulosa cells do not reveal euploid polyploidization in spontaneous or induced ovulatory cycles, the detected cells with increased DNA content should be interpreted as aneuploid, i.e. with chromosomal aberrations, and so their presence could also be discussed in connection with the hypothetical risk of prospective neoplastic transformation of the tissue.

Adult↗
PubMed1996

Three-dimensional confocal laser scanning DNA ploidy cytometry in thick histological sections.

DNA ploidy measurement by flow (FCM) or image cytometry (ICM) of single cell suspensions of solid tumour has prognostic value, but it would be a definite advantage if the assessment could be done on histological sections. However, this is usually not possible by means of standard ICM, due to the capping of nuclei in thin sections, or overlap in thick sections. Three-dimensional (3D) microscopy by means of confocal laser scanning microscopy (CLSM) could solve this problem in theory but the results published so far are not very satisfactory. A new method has been developed in which the DNA content of haploid (human testis spermatozoa), diploid, tetraploid, octaploid (human and rat liver and human spermatogonia), and near-triploid (human breast cancer) nuclei stained with YOYO-1 iodide has been measured by a newly developed 3D image cytometry method (3DICM) in 20 microns thick histological sections. YOYO-1 iodide is a new highly sensitive, specific, stoichiometric, and stable fluorescent dye for DNA. DNA ploidy of a breast cancer which was near-triploid with FCM and ICM was also assessed with 3DICM in a tissue section adjacent to the section used for FCM and ICM and the results were compared. The integrated 3DICM fluorescence intensity showed good linearity (r = 0.99) with the real DNA content of all nuclei analysed. In human tissue, the coefficient of variation of 3DICM for haploid (n = 12), diploid (n = 63), triploid (n = 13), tetraploid (n = 12), and octaploid (n = 3) ploidy distributions was 5.1, 6.6, 4.2, 4.0, and 0.6 per cent, respectively (n = the number of nuclei). For the rat liver, the CV of the diploid (n = 21), tetraploid (n = 31), and octaploid (n = 3) peaks was 6.7, 4.8, and 1.6 per cent, respectively. Repeated "blind' measurements of nuclei with different DNA indices showed excellent reproducibility between different observers (r = 0.98). It is concluded that the 3DICM method used is accurate, reproducible, and clinically feasible in thick histological sections. This is especially important in small lesions, or if the results of DNA ploidy measurement of single cell suspensions (by FCM) or imprints (by ICM) are inadequate.

Animals↗
PubMed1994

Large-cell acanthoma of the skin. A study by image analysis cytometry and immunohistochemistry.

Although large-cell acanthoma is a well-known clinicopathological entity, its biologic spectrum and nature are still subject to debate. We studied seven cases of large-cell acanthoma by image analysis cytometry for DNA content and by immunohistochemistry, using antibodies to proliferating cell nuclear antigen (PCNA)/cyclin. The data were compared with individual cases of seborrheic keratosis (SK), actinic keratosis (AK), and Bowen's disease (BD). The DNA distribution of large-cell acanthoma was variable. There were varying peaks at the DNA index values of 1 and 2 (diploid and tetraploid values), but all cases contained a significant aneuploid population between DNA index of 1 and 2. The mean DNA index was 1.44 (1.27-1.77); 1-20% of the cells exceeded 2, and 0-2% exceeded 3. The DNA index for lesions in the other differential diagnostic groups studied was as follows: SK, 1.0; AK, 1.4; BD, 1.8. The percentage of cells with positive nuclear staining for PCNA/cyclin was < 20% in all cases of large-cell acanthoma. The discrepancy between the high number of aneuploid and tetraploid cells observed on the DNA distribution curve and the lack of evidence for significant proliferation based on immunohistochemical stains suggest that these cells are resting cells with abnormal DNA clone. Although these results provide additional information about the biologic nature of large-cell acanthoma, they do not resolve the controversial nosologic status of lesions in this histologic group.

DNA↗
PubMed1999

Image analysis software for automatic DNA ploidy assessment of archival solid tumours.

BACKGROUND: Image cytometry has proved to provide a good alternative to flow cytometry for DNA ploidy measurement of archival tumors. However, when interactively done this technique is unable to give statistically valuable results within an acceptable time for clinical oncology. METHODS: An image cytometer was developed for fully automatic DNA ploidy quantitation, focusing efforts on speed and accuracy. Software functionalities include systematic acquisition of fields on a microscopic slide, detection, localization and sorting of nuclei, computation of the DNA content together with post-processing tools, for a deeper analysis of the DNA ploidy diagram. RESULTS: DNA ploidy analysis of archival breast carcinoma samples illustrates the accuracy of DNA ploidy measurements and the sensitivity in the detection of DNA ploidy abnormalities as a result of cell sorting. CONCLUSIONS: Fully automatic image cytometry is able to combine qualities of flow cytometry (automatic analysis of a statistically significant collection of cell nuclei) with additional advantages: sorting of unwanted events (debris, stromal and inflammatory cell nuclei) and facilities for an a posteriori control of the quality of cell selection. This method is well suited to DNA ploidy analysis of archival cancer samples.

Aneuploidy↗
PubMed2006

Prognostic impact of Deoxyribonucleic acid (DNA) image analysis cytometry and immunohistochemical expression of Ki67 in surgically resected non-small cell lung cancers.

BACKGROUND: The aim of the present study was to evaluate the prognostic significance of DNA ploidy and Ki67 expression in non-small cell lung carcinoma (NSCLC). METHODS: This prospective study included 96 patients with stages I-IIIA NSCLC who underwent surgical excision. DNA image analysis cytometry was applied on imprints. Calculation of the DNA index (DI) and the 5c exceeding rate (5cER) was performed and the histograms were classified as peridiploid, peritetraploid, and x-ploid-multiploid. The Ki67 immunoreactivity was determined according to the avidin-biotin complex immunoperoxidase method. RESULTS: DNA histogram classification disclosed 30 peridiploid cases, 15 peritetraploid and 51 x-ploid-multiploid. Forty-eight cases (50%) had 5cER > 5%. The Ki67 immunoreactivity was below 25% in 53 tumors (62.4%) and above 25% in 32 (32.6%). Our results revealed the existence of a statistically significant relationship of DNA ploidy with nodal status (p = 0.042) and grade (p = 0.005). Adenocarcinomas and large cell carcinomas were more frequently encountered in x-ploid-multiploid tumors as compared to squamous cell carcinomas, which were more frequently peridiploid (p = 0.003). 5cER showed statistically significant association with nodal status (p = 0.037). Univariate analysis with respect to survival revealed significant association with stage (p < 0.001), nodal status (p < 0.001), tumor status (p < 0.001), DNA ploidy (p = 0.008) and 5cER (p = 0.0124). Multivariate analysis revealed stage and ploidy status as independent factors: peridiploid tumors were associated with better survival as compared to x-ploid-multiploid tumors (p = 0.022). CONCLUSION: Our results suggest that DNA ploidy, as determined by image analysis, provides an independent prognostic parameter for patients with NSCLC and thus, could be used to identify a subset of patients with more aggressive tumors.

Adult↗
PubMed1999

Discrimination between benign and malignant melanocytic skin lesions by multivariate analysis, quantitative S-100 immunohistochemistry, nuclear morphometry and DNA cytometry.

OBJECTIVE: To determine whether combined quantitative immunohistochemistry of S-100, nuclear morphometry and DNA image cytometry improves discrimination between benign and malignant melanocytic skin lesions (MSLs). STUDY DESIGN: S-100 protein expression was measured in tissue sections of MSLs using an image cytometry system. Localized areas of high S-100 expression were used to identify regions in sequential, facing sections in which morphometric and cytometric features of nuclei, including DNA ploidy, were also measured. RESULTS: Malignant cases had significantly higher S-100 protein staining intensity, larger nuclei and greater DNA content (P < .05). High staining intensity for S-100 protein weakly correlated with variation in size of the mean nuclear area (P = .04) and DNA content (P = .03). Combining the features of nuclear area and DNA integrated optical density in areas of high-intensity staining for S-100 protein discriminated more accurately between 12 benign and 16 malignant areas than any of the features along (P = .0003). CONCLUSION: Combined multivariate quantitative immunohistochemical, morphometric and DNA cytometric analysis greatly improves discrimination between benign MSLs and malignant melanoma. Larger test sets are required to confirm the promising results of this initial study.

Cell Nucleus↗