Automated identification of diploid reference cells in cervical smears using image analysis.

BACKGROUND Acquisition of DNA ploidy histograms by image analysis may yield important information regarding the behavior of premalignant cervical lesions. Accurate selection of nuclei for DNA measurement is an important prerequisite for obtaining reliable data. Traditionally, manual selection of nuclei of diagnostic and reference cells is performed by an experienced cytotechnologist. In the present study, a method for the fully automated identification of nuclei of diploid epithelial reference cells in Feulgen- restained Papanicolaou (PAP) smears is described. METHODS The automated procedure consists of a decision tree implemented on the measurement device, containing nodes with feature threshold values and multivariate discriminant functions. Nodes were constructed to recognize debris and inflammatory cells, as well as diploid and nondiploid epithelial cells of the uterine cervix. Evaluation of the classifier was performed by comparing resulting diploid integrated optical densities with those from manually selected reference cells. RESULTS AND CONCLUSION On average, automatically acquired values deviated 2.4% from manually acquired values, indicating that the method described in this paper may be useful in cytometric practice.

[1]  D. Wittekind,et al.  Standardization of the Feulgen-Schiff technique , 2004, Histochemistry.

[2]  D. K. Das,et al.  Microphotometric DNA analysis in moderate dysplasia of the uterine cervix. Correlation to the progression and regression of the lesion. , 1990, Acta oncologica.

[3]  S Farrow,et al.  Semi-automated cervical smear pre-screening systems: an evaluation of the Cytoscan-110. , 1993, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[4]  R. Kiss,et al.  Modification of tumor ploidy level via the choice of tissue taken as diploid reference in the digital cell image analysis of Feulgen-stained nuclei. , 1992, Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc.

[5]  Keerti V. Shah,et al.  Genital warts and cervical cancer , 1984 .

[6]  F Giroud,et al.  Standardisation of DNA quantitation by image analysis: quality control of instrumentation. , 1999, Cytometry.

[7]  Pasquale Chieco,et al.  A user's guide for avoiding errors in absorbance image cytometry: a review with original experimental observations , 2004, The Histochemical Journal.

[8]  M Aubele,et al.  Interactive cytometry, chance or evil of bias? , 1992, Pathology, research and practice.

[9]  H. Sachs,et al.  [Cytofotometrically Determined DNA-pattern of premalignant cases of dysplasia of the cervix uteri. Contribution to the problem of morphologic definition and clinical sigificance of these lesions (author's transl)]. , 1975, Archiv fur Gynakologie.

[10]  E K Schulte,et al.  Standardization of the Feulgen reaction: the influence of chromatin condensation on the kinetics of acid hydrolysis. , 1990, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[11]  I. Ellis,et al.  Quality assurance in DNA image analysis on diploid cells. , 1997, Cytometry.

[12]  B Palcic,et al.  The use of an automated image cytometer for screening and quantitative assessment of cervical lesions in the British Columbia Cervical Smear Screening Programme , 1997, Cytopathology : official journal of the British Society for Clinical Cytology.

[13]  A. F. M. Moorman,et al.  Basic strategies for valid cytometry using image analysis , 1997, The Histochemical Journal.

[14]  R. Kiss,et al.  Characterization of factors in routine laboratory protocols that significantly influence the Feulgen reaction. , 1993, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[15]  B Palcic,et al.  Nuclear texture measurements in image cytometry. , 1995, Pathologica.

[16]  A Marchevsky,et al.  Quality assurance issues in DNA image cytometry. , 1996, Cytometry.

[17]  K Rodenacker,et al.  Problems concerning the quality of DNA measurements on Feulgen-stained imprints. A study of five fixation techniques. , 1994, Analytical and quantitative cytology and histology.

[18]  J. A. van der Laak,et al.  Development and validation of a computerized cytomorphometric method to assess the maturation of vaginal epithelial cells. , 1999, Cytometry.

[19]  P Herlin,et al.  Image analysis software for automatic DNA ploidy assessment of archival solid tumours. , 1999, Cytometry.

[20]  L. Koss The Papanicolaou test for cervical cancer detection. A triumph and a tragedy. , 1989, JAMA.

[21]  Calum MacAulay,et al.  DNA-Cytometry of Progressive and Regressive Cervical Intraepithelial Neoplasia , 1998, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[22]  B Palcic,et al.  Comparison of three different methods for automated classification of cervical cells. , 1992, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[23]  J H Tucker,et al.  Interval-coded texture features for artifact rejection in automated cervical cytology. , 1988, Cytometry.

[24]  J. Baak,et al.  Quantitative microscopical and confocal laser scanning microscopy for intermediate endpoint biomarkers in breast cancer: Potential and reproducibility , 1993, Journal of cellular biochemistry. Supplement.

[25]  I O Ellis,et al.  Interlaboratory comparison of DNA image analysis. , 1996, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[26]  G. Haroske,et al.  Part I: Basic Considerations and Recommendations for Preparation, Measurement and Interpretation , 1998, Analytical Cellular Pathology : the Journal of the European Society for Analytical Cellular Pathology.

[27]  Fritz Albregtsen,et al.  A Review of Caveats in Statistical Nuclear Image Analysis , 1998, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[28]  M. Us-Krasovec,et al.  The Effect of Primary Fixation with Standard Postfixation and the Duration of Hydrolysis on Nuclear Features in Image Cytometry , 1998, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[29]  A. Vinogradov Chromatin signal in genome size measurements. , 1999, Cytometry.

[30]  W. Auffermann,et al.  DNA-cytometric diagnosis of prospective malignancy in borderline lesions of the uterine cervix. , 1986, Acta cytologica.

[31]  B Palcic,et al.  Differences in quantitative nuclear features between ductal carcinoma in situ (DCIS) with and without accompanying invasive carcinoma in the surrounding breast. , 1995, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.