Analytical Cellular Pathology - Volume 26, issue 3

Authors: Ploeger, Lennert S. | Beliën, Jeroen A.M. | Poulin, Neal M. | Grizzle, William | Diest, Paul J. van

Article Type: Research Article

Abstract: Background: Confocal Laser Scanning Microscopy (CLSM) provides the opportunity to perform 3D DNA content measurements on intact cells in thick histological sections. So far, sample size has been limited by the time consuming nature of the technology. Since the power of DNA histograms to resolve different stemlines depends on both the sample size and the coefficient of variation (CV) of histogram peaks, interpretation of 3D CLSM DNA histograms might be hampered by both a small sample size and a large CV. The aim of this study was to analyze the required CV for 3D CLSM DNA histograms given a realistic …sample size. Methods: By computer simulation, virtual histograms were composed for sample sizes of 20000, 10000, 5000, 1000, and 273 cells and CVs of 30, 25, 20, 15, 10 and 5%. By visual inspection, the histogram quality with respect to resolution of G0/1 and G2/M peaks of a diploid stemline was assessed. Results: As expected, the interpretability of DNA histograms deteriorated with decreasing sample sizes and higher CVs. For CVs of 15% and lower, a clearly bimodal peak pattern with well distinguishable G0/1 and G2/M peaks were still seen at a sample size of 273 cells, which is our current average sample size with 3D CLSM DNA cytometry. Conclusions: For unambiguous interpretation of DNA histograms obtained using 3D CLSM, a CV of at most 15% is tolerable at currently achievable sample sizes. To resolve smaller near diploid stemlines, a CV of 10% or better should be aimed at. With currently available 3D imaging technology, this CV is achievable. Show more

Keywords: Confocal laser scanning microscopy, DNA ploidy, histogram quality, sample size, computer simulation

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 93-99, 2004

Authors: Chiu, David | Guillaud, Martial | Cox, Dennis | Follen, Michele | MacAulay, Calum

Article Type: Research Article

Abstract: Aims: Optical technologies have shown some promise for improving the care of cervical neoplasia. We are currently evaluating fluorescence and reflectance spectroscopy and quantitative cyto‐histopathology for cervical neoplasia screening and diagnosis. Here we describe the establishment and application of a quality assurance (QA) system for detecting system malfunctions and assessing the comparability of four image cytometers used in a multicenter clinical trial. Methods: Our QA system involves three levels of evaluation based on the periodicity and complexity of the measurements. We implemented our QA system at three image cytometers at the British Columbia Cancer Agency and one at M.D. …Anderson Cancer Center. The measurements or tasks were performed daily, monthly, and semi‐annually. The current and voltage of the lamp, the calibration image characteristics, and the room temperature were checked daily. Long‐term stability over time, short‐term variability over time, and spatial response field uniformity were evaluated monthly. Camera linearity was measured semi‐annually. Control charts based on statistical process control techniques were used to detect when the system did not perform optimally. Results: Daily measurements have shown good consistency in room temperature, lamp and calibration behaviour. Monthly measurements have shown small coefficients of variation between and within the four devices. There have been greater differences between sessions than within sessions. Comparability among the four systems is reasonably good. Semi‐annual measurements have shown stable camera linearity. QA events were detected using the QA system. Multiple examples of event detection leading to correction of system malfunction are described in this report. Conclusions: QA programs are critical for ensuring data integrity and therefore for the conduct of multicenter clinical trials. Show more

Keywords: Image cytometry, quality assurance, quality control, statistical process control, histopathology, quantitative, cytology, quantitative, technology assessment, cervical intraepithelial neoplasia, lesions, squamous intraepithelial

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 101-117, 2004

Authors: Hausmann, Michael | Cremer, Christoph | Linares‐Cruz, Gustavo | Nebe, Thomas C. | Peters, Katharina | Plesch, Andreas | Tham, Jochen | Vetter, Michael | Werner, Martin

Article Type: Research Article

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 119-124, 2004

Authors: Gerger, Armin | Bergthaler, Patrick | Smolle, Josef

Article Type: Research Article

Abstract: Aims. In tissue counter analysis (TCA) digital images of complex histologic sections are dissected into elements of equal size and shape, and digital information comprising grey level, colour and texture features is calculated for each element. In this study we assessed the feasibility of TCA for the quantitative description of amount and also of distribution of immunostained material. Methods. In a first step, our system was trained for differentiating between background and tissue on the one hand and between immunopositive and so‐called other tissue on the other. In a second step, immunostained slides were automatically screened and the procedure …was tested for the quantitative description of amount of cytokeratin (CK) and leukocyte common antigen (LCA) immunopositive structures. Additionally, fractal analysis was applied to all cases describing the architectural distribution of immunostained material. Results. The procedure yielded reproducible assessments of the relative amounts of immunopositive tissue components when the number and percentage of CK and LCA stained structures was assessed. Furthermore, a reliable classification of immunopositive patterns was found by means of fractal dimensionality. Conclusions. Tissue counter analysis combined with classification trees and fractal analysis is a fully automated and reproducible approach for the quantitative description in immunohistology. Show more

Keywords: Image analysis, tissue counter analysis, quantification, fractal geometry, immunostaining

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 125-134, 2004

Authors: Kruse, Arnold‐Jan | Buhr‐Wildhagen, Susanne | Janssen, Emiel A. | Baak, Jan P.A.

Article Type: Research Article

Abstract: Aim: To assess the correlation between syntactic structure analysis (SSA) features, revised dysplasia grade and the presence of high‐risk human papillomavirus DNA in cervical intraepithelial neoplasia (CIN). Materials and methods: HPV polymerase chain reaction (PCR) was assessed in 101 consecutive biopsies and consensus in CIN grade between the experts occurred in 88 cases (CIN1=16, CIN2=27, CIN3=45). SSA was performed in the diagnostic histological section of the CIN lesions in these patients and SSA features were compared with the blind review CIN grade, and presence/absence of high‐risk HPV DNA. Results: One of the SSA features (points from which the surrounding surfaces …has 4 edges, PECO‐4) was significantly different between all three consensus CIN grades. Many more features revealed significant differences between CIN1 and CIN2 or between CIN2 and CIN3 cases. With stepwise discriminant analysis, the best multivariate combination of features to distinguish the different CIN grades were the Maximum MST Line Length (MML) and the Area Disorder. Crude overall classification of the consensus grades with these features was 69%. The MML and the Area Disorder is also the best combination to distinguish cases with and without high‐risk HPV DNA (77% correct classifications). Conclusions: SSA features are correlated with both CIN grade and presence of high‐risk HPV DNA, but the discrimination power is not good enough to be used as a routine method for quality control of subjective grade or as a surrogate marker for high‐risk HPV DNA presence. Show more

Keywords: Cervix, dysplasia, CIN, HPV, SSA, differentiation

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 135-141, 2004

Authors: Nguyen, Vu Quoc Huy | Grote, Hans Jürgen | Pomjanski, Natalia | Knops, Kristiane | Böcking, Alfred

Article Type: Research Article

Abstract: In the present study, the aim has been to investigate the interobserver reproducibility of DNA‐image‐cytometry (DNA‐ICM) applied to routine Pap smears classified as Atypical Squamous Cells of Undetermined Significance (ASCUS) or higher lesions (ASCUS+). 202 Pap smears diagnosed as ASCUS or higher were included in the study. After cytological assessment, smears underwent restaining according to Feulgen. First measurements were performed as routine workup. The second measurements were blinded to the result of the first and consecutively performed. DNA‐ICM met the consensus statements of the European Society of Analytical Cellular Pathology (ESACP). Interobserver agreement was assessed by calculating Kappa statistics. The …diagnosis of DNA‐aneuploidy in the first measurements was confirmed in all cases. Second measurement detected 12 additional cases with aneuploidy. Nine out of these cases were classified as aneuploidy by detection of 9c Exceeding Events (9cEE). In three cases stemline‐aneuploidy was disclosed. The overall proportion of observed agreement was 94.1%, κ=0.87, 95% CI=0.74–0.99. Our study shows a good interobserver reproducibility of DNA‐ICM performed on cervical smears with ASCUS or higher lesions. DNA‐ICM thus represents a highly reproducible diagnostic procedure. Show more

Keywords: Interobserver reproducibility, DNA‐image‐cytometry, cervical cytology

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 143-150, 2004

Article Type: Abstract

Citation: Analytical Cellular Pathology, vol. 26, no. 3, pp. 151-165, 2004