Analytical Cellular Pathology - Volume 25, issue 2

Authors: Gerstner, Andreas O.H. | Machlitt, Julia | Welkoborsky, Hans‐Jürgen | Bootz, Friedrich | Tárnok, Attila

Article Type: Research Article

Abstract: Aim: To test laser scanning cytometry (LSC) for the analysis of ploidy in squamous cell carcinoma of the hypopharynx (SCCH) and to develop a routine application for minimal samples such as fine needle aspirate biopsies (FNABs). Methods: From 11 individuals 30 FNABs of primary tumors (n=11) and lymphatic metastases of SCCH (n=11) and non‐metastatic lymph nodes (n=8) are analyzed by LSC. This microscope based instrument scans the cells after immobilization on a glass slide and after double staining of cytokeratin and DNA. The location of each cell is stored with the fluorescence data. Therefore the morphology of every cell can …be documented by re‐staining with H&E and re‐localization on the slide. Additionally, aliquots are Feulgen‐stained for image cytometry in 8 specimens. Results: The diploid reference peak is identified taking leukocytes as internal standard. The DNA‐index of the carcinoma cells ranges from 0.4 to 3.8. Comparison with image cytometry shows good correlation (r=0.89). Conclusion: LSC provides a reliable and objective way to determine the ploidy of SCCH pre‐operatively. Colour figures can be viewed on http://www.esacp.org/acp/2003/25‐2/gerstner.htm. Show more

Keywords: Ploidy, carcinoma of the hypopharynx, comparative study, LSC, slide‐based cytometry, image cytometry, Feulgen‐staining, eosinophils

Citation: Analytical Cellular Pathology, vol. 25, no. 2, pp. 51-62, 2003

Authors: Kim, Nga Tran | Elie, Nicolas | Plancoulaine, Benoît | Herlin, Paulette | Coster, Michel

Article Type: Research Article

Abstract: Relative abundance of tumour angiogenesis has been shown to be of clinical relevance in cancers of various locations such as the ovary. Nevertheless, several problems are encountered when quantifying tumour microvessels: (i) as many other tumour markers, vascularity pattern is often heterogeneous within the tumour mass and even within the same histological section. As a consequence, an adequate acquisition method must be developed for accurate field sampling. (ii) Manual microvessel counting is long, tedious and subject to poor reproducibility. Introduction in routine practice requires a fast, reproducible and reliable automatic image processing. In this study we present an original …procedure combining a slide scanner image acquisition and a fully automatic image analysis sequence. The slide scanner offers the advantage of recording an image of the whole histological section for subsequent automatic blood vessel detection and hot spot area location. Microvessel density and surface fraction were measured for the whole section as well as within hot spots. Different immunostaining methods were tested in order to optimise the procedure. Moreover, the method proposed was submitted to a quality control procedure, with reference to interactive identification of microvessels at scanner level. This experiment showed that 93 to 97% of blood vessels were detected, according to the staining protocol used. Colour figures can be viewed on http://www.esacp.org/acp/2003/25‐2/kim.htm. Show more

Keywords: Angiogenesis, quantification, image analysis, slide scanner

Citation: Analytical Cellular Pathology, vol. 25, no. 2, pp. 63-75, 2003

Authors: Kristiansen, Glen | Yu, Yongwei | Schlüns, Karsten | Sers, Christine | Dietel, Manfred | Petersen, Iver

Article Type: Research Article

Abstract: Expression of MCAM is observed in a variety of human malignancies. We aimed to determine the rate of MCAM expression in our non‐small cell lung cancer (NSCLC) collection and to clarify its correlation with clinicopathological parameters and patient survival. 85 NSCLC were analysed immunohistochemically using a monoclonal MCAM antibody (clone N1238) on an NSCLC tissue micro array. The staining was semiquantitatively scored. We found MCAM expression in 51% of NSCLC, preferentially squamous cell carcinomas (p=0.004). No other correlations to tumour size, grade, or stage were found. Univariate survival analysis showed no significant differences of MCAM positive and negative tumours. In …adenocarcinomas however, MCAM positivity was significantly associated with shorter patient survival (p=0.016). We conclude, that MCAM is expressed in a high proportion of NSCLC and might be predictive of shortened patient survival in adenocarcinomas of the lung. Colour figure can be viewed on http://www.esacp.org/acp/2003/25‐2/kristiansen.htm Show more

Keywords: Non‐small cell lung cancer, MCAM, CD146, multiple tissue array, immunohistochemistry

Citation: Analytical Cellular Pathology, vol. 25, no. 2, pp. 77-81, 2003

Authors: Grote, Hans Jürgen | Schmiemann, Viola | Sarbia, Mario | Böcking, Alfred

Article Type: Research Article

Abstract: Objective: To date, there are only few systematic reports on the quality of DNA extracted from routine diagnostic cytologic specimens. It was the aim of the present study to evaluate the ability of 50% ethanol/2% carbowax (Saccomanno fixative) to preserve bronchial secretions with high quality genomic DNA as well as to compare different DNA extraction methods. Methods: DNA was extracted from 45 bronchial aspirates by four different extraction protocols. Beside DNA yield, DNA quality with regard to purity, integrity, and PCR success rate were investigated. Results: No fragmentation of sample DNA due to the fixative was detected. It …was preserved as high molecular weight DNA. DNA yield, purity, and integrity were dependent on the DNA extraction method to some extend. Irrespective of the DNA extraction method the PCR success rate for amplification of β‐globin gene fragments (268, 536, and 989 bp) was 100%. Conclusions: A fixative containing 50% ethanol/2% carbowax preserves high quality DNA which is well suited for PCR‐based assays regardless of the extraction protocol used. The selection of the DNA extraction protocol has to be adjusted to the circumstances of application. Show more

Keywords: Molecular cytology, fixative, PCR, bronchi, DNA extraction

Citation: Analytical Cellular Pathology, vol. 25, no. 2, pp. 83-88, 2003

Authors: Jee, Kowan J. | Gong, Gyungyub | Ahn, Sei Hyun | Park, Jeong Mi | Knuutila, Sakari

Article Type: Research Article

Abstract: We studied DNA copy number changes by CGH and allelic imbalance (AI) on 3p by LOH analysis on 22 phyllodes tumours (PT) of the breast in order to gain insight into the genetic basis of tumour progression in PT. Copy number changes were observed in 14 cases (63%). Gain in 1q with 1q21–23 as the minimal overlapping area was seen in 12 cases (55%). The gain was observed both in benign and malignant tumours. Our study did not reveal any DNA copy number changes or allelic loss on 3p. The results suggest that DNA copy number changes are not associated …with the histological grade or clinical behaviour of PT and the chromosomal changes on 3p appear to be rare. Colour figure can be viewed on http://www.esacp.org/acp/2003/25‐2/jee.htm Show more

Keywords: Phyllodes tumour, comparative genomic hybridization, allelic loss on 3p, chromosome 1q21–q23

Citation: Analytical Cellular Pathology, vol. 25, no. 2, pp. 89-93, 2003

Authors: Sun, Xiao Rong | Zheng, Yonghong | MacAulay, Calum | Lam, Stephen | Doudkine, Alexei | Palcic, Branko

Article Type: Research Article

Abstract: Malignancy associated changes (MAC) can be defined as subtle morphological and physiologic changes that are found in ostensibly normal cells of patients harboring malignant disease. It has been postulated that MAC have a potential to become a useful tool in detection, diagnosis and prognosis of malignant diseases. An in vitro cell culture model system was designed to study interactions between non‐small cell lung cancer (NSCLC) and the normal bronchial epithelium of the human respiratory tract in vivo to see if the MAC‐like phenomenon can be detected in such a system. In this study we examined changes in nuclear features of …normal human bronchial epithelial cells (NHBE) when they were co‐cultured with cells derived from a lung cancer cell line NCI‐H460. Using discriminant function analysis, nuclear features were determined which allow maximal discrimination between normal cells incubated with or without cancerous cells. Our results demonstrate that MAC appear to be specific to changes induced by malignancy, and that these changes differ from those induced by growth factors in the serum. This study provides evidence in support to the hypothesis that MAC are induced by a soluble factor(s) released by malignant cells. Colour figure can be viewed on http://www.esacp.org/acp/2003/25‐2/sun.htm Show more

Keywords: Cell co‐culture, image cytometry, nuclear features, malignancy associated changes

Citation: Analytical Cellular Pathology, vol. 25, no. 2, pp. 95-102, 2003