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Evaluation of colon cancer histomorphology: a comparison between formalin and PAXgene tissue fixation by an international ring trial

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Abstract

The aim of our study was to evaluate the quality of histo- and cytomorphological features of PAXgene-fixed specimens and their suitability for histomorphological classification in comparison to standard formalin fixation. Fifteen colon cancer tissues were collected, divided into two mirrored samples and either formalin fixed (FFPE) or PAXgene fixed (PFPE) before paraffin embedding. HE- and PAS-stained sections were scanned and evaluated in a blinded, randomised ring trial by 20 pathologists from Europe and the USA using virtual microscopy. The pathologists evaluated histological grading, histological subtype, presence of adenoma, presence of lymphovascular invasion, quality of histomorphology and quality of nuclear features. Statistical analysis revealed that the reproducibility with regard to grading between both fixation methods was rather satisfactory (weighted kappa statistic (k w) = 0.73 (95 % confidence interval (CI), 0.41–0.94)), with a higher agreement between the reference evaluation and the PFPE samples (k w = 0.86 (95 % CI, 0.67–1.00)). Independent from preservation method, inter-observer reproducibility was not completely satisfactory (k w = 0.60). Histomorphological quality parameters were scored equal or better for PFPE than for FFPE samples. For example, overall quality and nuclear features, especially the detection of mitosis, were judged significantly better for PFPE cases. By contrast, significant retraction artefacts were observed more frequently in PFPE samples. In conclusion, our findings suggest that the PAXgene Tissue System leads to excellent preservation of histomorphology and nuclear features of colon cancer tissue and allows routine morphological diagnosis.

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References

  1. Agresti A (ed) (1990) Categorical data analysis. John Wiley and Sons, New York

    Google Scholar 

  2. Barresi V, Reggiani Bonetti L, Branca G et al (2012) Colorectal carcinoma grading by quantifying poorly differentiated cell clusters is more reproducible and provides more robust prognostic information than conventional grading. Virchows Arch 461:621–628

    Article  PubMed  Google Scholar 

  3. Bosetti C, Mclaughlin JK, Tarone RE et al (2008) Formaldehyde and cancer risk: a quantitative review of cohort studies through 2006. Ann Oncol Off J Eur Soc Med Oncol ESMO 19:29–43

    Article  CAS  Google Scholar 

  4. Bosman FT, Carneiro F, Hruban RH et al (eds) (2010) WHO classification of tumours of the digestive system. International Agency for Research on Cancer (I A R C)

  5. Chandler I, Houlston RS (2008) Interobserver agreement in grading of colorectal cancers—findings from a nationwide Web-based survey of histopathologists. Histopathology 52:494–499

    Article  PubMed  CAS  Google Scholar 

  6. Chetty R, Gill P, Govender D et al (2012) International study group on rectal cancer regression grading: interobserver variability with commonly used regression grading systems. Hum Pathol 43:1917–1923

    Article  PubMed  Google Scholar 

  7. Corletto V, Verderio P, Giardini R et al (1998) Evaluation of residual cellularity and proliferation on preoperatively treated breast cancer: a comparison between image analysis and light microscopy analysis. Anal Cell Pathol 16:83–93

    PubMed  CAS  Google Scholar 

  8. Ergin B, Meding S, Langer R et al (2010) Proteomic analysis of PAXgene-fixed tissues. J Proteome Res 9:5188–5196

    Article  PubMed  CAS  Google Scholar 

  9. Fleiss JL, Levin B, Paik MC (eds) (1981) Statistical methods for rates and proportions. John Wiley and Sons, New York

    Google Scholar 

  10. Groelz D, Sobin L, Branton P et al (2013) Non-formalin fixative versus formalin-fixed tissue: a comparison of histology and RNA quality. Exp Mol Pathol 94:188–194

    Article  PubMed  CAS  Google Scholar 

  11. Gündisch S, Schott C, Wolff C et al (2013) The PAXgene((R)) tissue system preserves phosphoproteins in human tissue specimens and enables comprehensive protein biomarker research. PLoS One 8:e60638

    Article  PubMed  PubMed Central  Google Scholar 

  12. Harris EI, Lewin DN, Wang HL et al (2008) Lymphovascular invasion in colorectal cancer: an interobserver variability study. Am J Surg Pathol 32:1816–1821

    Article  PubMed  PubMed Central  Google Scholar 

  13. Holman CD (1984) Analysis of interobserver variation on a programmable calculator. Am J Epidemiol 120:154–160

    PubMed  CAS  Google Scholar 

  14. Italian Network for Quality Assurance of Tumour Biomarkers G (2005) Quality control for histological grading in breast cancer: an Italian experience. Pathologica 97:1–6

    Google Scholar 

  15. Jemal A, Center MM, Desantis C et al (2010) Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomarkers Prev 19:1893–1907

    Article  PubMed  Google Scholar 

  16. Jemal A, Siegel R, Xu J et al (2010) Cancer statistics, 2010. CA Cancer J Clin 60:277–300

    Article  PubMed  Google Scholar 

  17. Kap M, Smedts F, Oosterhuis W et al (2011) Histological assessment of PAXgene tissue fixation and stabilization reagents. PLoS One 6:e27704

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  18. Kayser K (2012) Introduction of virtual microscopy in routine surgical pathology—a hypothesis and personal view from Europe. Diagn Pathol 7:48

    Article  PubMed  PubMed Central  Google Scholar 

  19. Kim S, Park HK, Jung HY et al (2013) ERG immunohistochemistry as an endothelial marker for assessing lymphovascular invasion. Korean J Pathol 47:355–364

    Article  PubMed  PubMed Central  Google Scholar 

  20. Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33:159–174

    Article  PubMed  CAS  Google Scholar 

  21. Nielsen GD, Wolkoff P (2010) Cancer effects of formaldehyde: a proposal for an indoor air guideline value. Arch Toxicol 84:423–446

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  22. O’donnell RK, Feldman M, Mick R et al (2008) Immunohistochemical method identifies lymphovascular invasion in a majority of oral squamous cell carcinomas and discriminates between blood and lymphatic vessel invasion. J Histochem Cytochem 56:803–810

    Article  PubMed  PubMed Central  Google Scholar 

  23. Oetjen J, Aichler M, Trede D et al (2013) MRI-compatible pipeline for three-dimensional MALDI imaging mass spectrometry using PAXgene fixation. J Proteomics 90:52–60

    Article  PubMed  CAS  Google Scholar 

  24. Paradiso A, Ellis IO, Zito FA et al (2009) Short- and long-term effects of a training session on pathologists’ performance: the INQAT experience for histological grading in breast cancer. J Clin Pathol 62:279–281

    Article  PubMed  CAS  Google Scholar 

  25. Shaw EC, Hanby AM, Wheeler K et al (2012) Observer agreement comparing the use of virtual slides with glass slides in the pathology review component of the POSH breast cancer cohort study. J Clin Pathol 65:403–408

    Article  PubMed  Google Scholar 

  26. Staff S, Kujala P, Karhu R et al (2013) Preservation of nucleic acids and tissue morphology in paraffin-embedded clinical samples: comparison of five molecular fixatives. J Clin Pathol 66:807–810

    Article  PubMed  Google Scholar 

  27. Thomas GD, Dixon MF, Smeeton NC et al (1983) Observer variation in the histological grading of rectal carcinoma. J Clin Pathol 36:385–391

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  28. Viertler C, Groelz D, Gündisch S et al (2012) A new technology for stabilization of biomolecules in tissues for combined histological and molecular analyses. J Mol Diagn 14:458–466

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was performed within the European consortium Standardisation and Improvement of Generic Pre-analytical Tools and Procedures for In Vitro Diagnostics (SPIDIA; www.spidia.eu), which is funded by the European Union within the Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 222916.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Pathology, Technische Universität München, Trogerstr. 18, 81675, Munich, Germany

    Sibylle Gündisch, Julia Slotta-Huspenina, Christina Schott, Enken Drecoll, Irene Esposito, Katja Specht, Gregor Babaryka, Heinz Höfler & Karl-Friedrich Becker

  2. Unit of Medical Statistics, Biometry and Bioinformatics, Fondazione IRCCS, Istituto Nazionale dei Tumori, via Venezian 1, 20133, Milan, Italy

    Paolo Verderio, Chiara Maura Ciniselli & Sara Pizzamiglio

  3. Institute of Pathology, Medical University of Graz, Auenbruggerplatz 25, 8036, Graz, Austria

    Christian Viertler, Kurt Zatloukal, Martin Asslaber & Koppany Bodó

  4. Department of Pathology, Erasmus Medical Center, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands

    Marcel Kap, Peter Riegman, Michael den Bakker & Jan den Hollander

  5. Institute of Pathology, Eberhard-Karls-University, Liebermeisterstr. 8, 72076, Tübingen, Germany

    Falko Fend

  6. Institute of Pathology, Ludwig-Maximilians-Universität München, Thalkirchner Str. 36, 80337, Munich, Germany

    Jens Neumann & Simone Reu

  7. Institute of Pathology, Universität Bern, Murtenstr. 31, Postfach 62, 3010, Bern, Switzerland

    Aurel Perren, Rupert Langer & Alessandro Lugli

  8. Pathology Rosenheim, Lilienweg 12, 83022, Rosenheim, Germany

    Ingrid Becker & Thomas Richter

  9. Department of Pathology, Ludwig-Aschoff-Haus, University Medical Center Freiburg, Breisacher Str. 115a, 79106, Freiburg, Germany

    Gian Kayser & Annette M. May

  10. IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, University of Porto, Rua Dr. Roberto Frias s/n, 4200-465, Porto, Portugal

    Fatima Carneiro & José Manuel Lopes

  11. Frederick National Laboratory for Cancer Research, The Cancer Human Biobank, National Cancer Institute, 6110 Executive Blvd, Suite 250, Rockville, MD, 20852, USA

    Leslie Sobin

  12. The SPIDIA Consortium http://www.spidia.eu

    Sibylle Gündisch, Paolo Verderio, Chiara Maura Ciniselli, Sara Pizzamiglio, Christina Schott, Christian Viertler, Kurt Zatloukal, Marcel Kap, Peter Riegman & Karl-Friedrich Becker

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  1. Sibylle Gündisch
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  2. Julia Slotta-Huspenina
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  12. Irene Esposito
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  16. Koppany Bodó
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  19. Falko Fend
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  20. Jens Neumann
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  26. Thomas Richter
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  27. Gian Kayser
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  29. Fatima Carneiro
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  30. José Manuel Lopes
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  31. Leslie Sobin
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Corresponding author

Correspondence to Karl-Friedrich Becker.

Additional information

Sibylle Gündisch and Julia Slotta-Huspenina contributed equally to this work.

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Gündisch, S., Slotta-Huspenina, J., Verderio, P. et al. Evaluation of colon cancer histomorphology: a comparison between formalin and PAXgene tissue fixation by an international ring trial. Virchows Arch 465, 509–519 (2014). https://doi.org/10.1007/s00428-014-1624-4

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  • DOI: https://doi.org/10.1007/s00428-014-1624-4

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