Cell and Tissue Research

, Volume 375, Issue 2, pp 371–381 | Cite as

Digital pathology imaging and computer-aided diagnostics as a novel tool for standardization of evaluation of aganglionic megacolon (Hirschsprung disease) histopathology

  • Florian Schilling
  • Carol E. Geppert
  • Johanna Strehl
  • Arndt Hartmann
  • Stefanie Kuerten
  • Axel Brehmer
  • Samir JabariEmail author
Regular Article


Based on a recently introduced immunohistochemical panel (Bachmann et al. 2015) for aganglionic megacolon (AM), also known as Hirschsprung disease, histopathological diagnosis, we evaluated whether the use of digital pathology and ‘machine learning’ could help to obtain a reliable diagnosis. Slides were obtained from 31 specimens of 27 patients immunohistochemically stained for MAP2, calretinin, S100β and GLUT1. Slides were digitized by whole slide scanning. We used a Definiens Developer Tissue Studios as software for analysis. We configured necessary parameters in combination with ‘machine learning’ to identify pathological aberrations. A significant difference between AM- and non-AM-affected tissues was found for calretinin (AM 0.55% vs. non-AM 1.44%) and MAP2 (AM 0.004% vs. non-AM 0.07%) staining measurements and software-based evaluations. In contrast, S100β and GLUT1 staining measurements and software-based evaluations showed no significant differences between AM- and non-AM-affected tissues. However, no difference was found in comparison of suction biopsies with resections. Applying machine learning via an ensemble voting classifier, we achieved an accuracy of 87.5% on the test set. Automated diagnosis of AM by applying digital pathology on immunohistochemical panels was successful for calretinin and MAP2, whereas S100β and GLUT1 were not effective in diagnosis. Our method suggests that software-based approaches are capable of diagnosing AM. Our future challenge will be the improvement of efficiency by reduction of the time-consuming need for large pre-labelled training data. With increasing technical improvement, especially in unsupervised training procedures, this method could be helpful in the future.


Calretinin Digital pathology Hirschsprung disease Immunohistochemistry Machine learning 



The present work was performed in fulfilment of the requirements of the Friedrich-Alexander Universität Erlangen-Nürnberg (FAU) for obtaining the degree ‘Dr. med.dent’ of Florian Schilling. We would like to thank Nicole Fuhrich and Definines Support for their excellent technical assistance.

Compliance with ethical standards

The use of the FFPE materials was approved by the local ethics committee (approval number: 85_12b, date 04/19/2012).

Conflict and interest statement

The authors declare that they have no conflict of interests.

Informed consent

Informed consent was obtained from all individual participants for whom identifying information is included in this article.

Supplementary material

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Florian Schilling
    • 1
    • 2
  • Carol E. Geppert
    • 2
  • Johanna Strehl
    • 2
  • Arndt Hartmann
    • 2
  • Stefanie Kuerten
    • 1
  • Axel Brehmer
    • 1
  • Samir Jabari
    • 1
    • 2
    Email author
  1. 1.Institute of Anatomy and Cell BiologyFriedrich-Alexander-Universität Erlangen-NürnbergErlangenGermany
  2. 2.Institute of PathologyFriedrich-Alexander-Universität Erlangen-NürnbergErlangenGermany

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