Analytical Cellular Pathology - Volume 15, issue 1

Authors: Choi, Heung‐Kook | Jarkrans, Torsten | Bengtsson, Ewert | Vasko, Janos | Wester, Kenneth | Malmström, Per‐Uno | Busch, Christer

Article Type: Research Article

Abstract: The possibility that computerized image analysis could increase the reproducibility of grading of bladder carcinoma as compared to conventional subjective grading made by pathologists was investigated. Object, texture and graph based analysis were carried out from Feulgen stained histological tissue sections. The object based features were extracted from gray scale images, binary images obtained by thresholding the nuclei and several other images derived through image processing operations. The textural features were based on the spatial gray‐tone co‐occurrence probability matrices and the graph based features were extracted from the minimum spanning trees connecting all nuclei. The large numbers of extracted features …were evaluated in relation to subjective grading and to factors related to prognosis using multivariate statistical methods and multilayer backpropagation neural networks. All the methods were originally developed and tested on material from one patient and then tested for reproducibility on entirely different patient material. The results indicate reasonably good reproducibility for the best sets of features. In addition, image analysis based grading showed almost identical correlation to mitotic density and expression of p53 protein as subjective grading. It should thus be possible to use this kind of image analysis as a prognostic tool for bladder carcinoma. Show more

Keywords: Image analysis, feature measurements, grading of bladder carcinoma, reproducibility

Citation: Analytical Cellular Pathology, vol. 15, no. 1, pp. 1-18, 1997

Authors: Rodenacker, Karsten | Aubele, Michaela | Hutzler, Peter | Umesh Adiga, P.S.

Article Type: Research Article

Abstract: In molecular pathology numerical chromosome aberrations have been found to be decisive for the prognosis of malignancy in tumours. The existence of such aberrations can be detected by interphase fluorescence in situ hybridization (FISH). The gain or loss of certain base sequences in the desoxyribonucleic acid (DNA) can be estimated by counting the number of FISH signals per cell nucleus. The quantitative evaluation of such events is a necessary condition for a prospective use in diagnostic pathology. To avoid occlusions of signals, the cell nucleus has to be analyzed in three dimensions. Confocal laser scanning microscopy is the means to …obtain series of optical thin sections from fluorescence stained or marked material to fulfill the conditions mentioned above. A graphical user interface (GUI) to a software package for display, inspection, count and (semi‐)automatic analysis of 3‐D images for pathologists is outlined including the underlying methods of 3‐D image interaction and segmentation developed. The preparative methods are briefly described. Main emphasis is given to the methodical questions of computer‐aided analysis of large 3‐D image data sets for pathologists. Several automated analysis steps can be performed for segmentation and succeeding quantification. However tumour material is in contrast to isolated or cultured cells even for visual inspection, a difficult material. For the present a fully automated digital image analysis of 3‐D data is not in sight. A semi‐automatic segmentation method is thus presented here. Show more

Keywords: Fluorescence in situ hybridization (FISH), confocal laser scanning microscopy, graphical user interface, 3‐D image analysis, segmentation

Citation: Analytical Cellular Pathology, vol. 15, no. 1, pp. 19-29, 1997

Authors: Leonardi, Elena | Cristofori, Antonella | Caffo, Orazio | Dalla Palma, Paolo

Article Type: Research Article

Abstract: The aim of the study was to correlate the DNA Index (DI) and S‐phase fraction (SPF) values determined by multiparametric flow cytometry in breast cancer (mainly T1 and T2 stages) with several clinico‐pathologic variables and other biological parameters. For this purpose, a total of 136 breast cancers were submitted to flow cytometry and to several types of immunohistochemical analyses. Among clinico‐pathologic data we considered pT, pN and grade and among immunohistochemical markers, hormonal receptors, P53, c‐erbB‐2 and MIB‐1. We found that DNA aneuploidy was strongly correlated with high tumoral grade, absence of hormonal receptors, high proliferation, as shown by high …MIB‐1 ({\scriptstyle\geq} 36%) and SPF values ({\scriptstyle\geq} 13.3%), and P53 positivity. Grouping the tumours according to their DI values, we observed a relative significantly higher correlation of the near‐triploid range carcinomas (DI 1.40–1.60) with immunohistochemical expression of P53 (p=0.0001 ). Near‐triploid DI was also associated with a high proliferative activity, expressed both by SPF (p=0.0001 ) and MIB‐1 reactivity (p=0.0001 ), high tumoral grade (p=0.0001 ) and presence of axillary metastases (p=0.03 ). These data suggest that DNA near‐triploid tumours in breast cancer may have a more aggressive behaviour in comparison with other DI classes. Show more

Keywords: Ploidy, breast cancer, P53, prognostic markers

Citation: Analytical Cellular Pathology, vol. 15, no. 1, pp. 31-45, 1997

Authors: Kronqvist, Pauliina | Kuopio, Teijo | Collan, Yrjö | Horvath, Csaba | Tamm, Ülle

Article Type: Research Article

Abstract: The intraobserver and interobserver reproducibility of computerized nuclear morphometry was determined in repeated measurements of 212 samples of invasive breast cancer. The influence of biological variation and the selection of the measurement area was also tested. Morphometrically determined mean nuclear profile area (Pearson’s r 0.89, grading efficiency (GE) 0.95) and standard deviation (SD) of nuclear profile area (Pearson’s r 0.84, GE 0.89) showed high reproducibility. In this respect, nuclear morphometry equals with other established methods of quantitative pathology and exceeds the results of subjective grading of nuclear atypia in invasive breast cancer. A training period of …eight days was sufficient to produce clear improvement in consistency of nuclear morphometry results. By estimating the sources of variation it could be shown that the variation associated with the measurement procedure itself is small. Instead, sample associated variation is responsible for the majority of variation in the measurements (82.9% in mean nuclear profile area and 65.9% in SD of nuclear profile area). This study points out that when standardized methods are applied computerized morphometry is a reproducible and reliable method of assessing nuclear atypia in invasive breast cancer. For further improvement special emphasize should be put on sampling rules of selecting the microscope fields and measurement areas. Show more

Keywords: Breast cancer, morphometry, nuclear profile area, reproducibility, nuclear pleomorphism, sources of variation

Citation: Analytical Cellular Pathology, vol. 15, no. 1, pp. 47-59, 1997