Semi-automated digital measurement as the method of choice for beta cell mass analysis

Violette Coppens; Gunter Leuckx; Yves Heremans; Willem Staels; Yannick Verdonck; Luc Baeyens; Nico De Leu; Harry Heimberg

doi:10.1371/journal.pone.0191249

Semi-automated digital measurement as the method of choice for beta cell mass analysis

PLoS One. 2018 Feb 6;13(2):e0191249. doi: 10.1371/journal.pone.0191249. eCollection 2018.

Authors

Violette Coppens^{1

2

3}, Gunter Leuckx¹, Yves Heremans¹, Willem Staels^{1

4}, Yannick Verdonck¹, Luc Baeyens¹, Nico De Leu^{1

5

6}, Harry Heimberg¹

Affiliations

¹ Beta cell Neogenesis, Vrije Universiteit Brussel, Brussels, Belgium.
² Collaborative Antwerp Psychiatric Research Institute, Faculty of Medicine and Health Sciences, University of Antwerp, Campus Drie Eiken, Antwerp, Belgium.
³ University Department of Psychiatry, Campus Duffel, Duffel, Belgium.
⁴ Department of Pediatrics, Division of Pediatric Endocrinology, Ghent University, Hospital and Department of Pediatrics and Genetics, Ghent, Belgium.
⁵ Department of Endocrinology, Universitair Ziekenhuis Brussel, Brussels, Belgium.
⁶ Department of Endocrinology, Algemeen Stedelijk Ziekenhuis Aalst, Aalst, Belgium.

Abstract

Pancreas injury by partial duct ligation (PDL) activates beta cell differentiation and proliferation in adult mouse pancreas but remains controversial regarding the anticipated increase in beta cell volume. Several reports unable to show beta cell volume augmentation in PDL pancreas used automated digital image analysis software. We hypothesized that fully automatic beta cell morphometry without manual micrograph artifact remediation introduces bias and therefore might be responsible for reported discrepancies and controversy. However, our present results prove that standard digital image processing with automatic thresholding is sufficiently robust albeit less sensitive and less adequate to demonstrate a significant increase in beta cell volume in PDL versus Sham-operated pancreas. We therefore conclude that other confounding factors such as quality of surgery, selection of samples based on relative abundance of the transcription factor Neurogenin 3 (Ngn3) and tissue processing give rise to inter-laboratory inconsistencies in beta cell volume quantification in PDL pancreas.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Automation*
Islets of Langerhans / pathology*

Grants and funding

This work was supported by the Instituut voor Innovatie door Wetenschap en Technologie (IWT) (project nr: 91113; URL: http://www.iwt.be/) provided a PhD scholarship for the first author (VC); Research Council, Vrije Universiteit Brussel (Strategic Research Project SRP35) to HH" "Belgian Science Policy (Interuniversity Attraction Pole phase VI, project P7/07 to HH; Fund for Scientific Research Flanders (FWO) (Research Grant G024512N) to HH; FWO-Odysseus GOF9116N) to LB; DON Foundation (www.sdon.nl) to HH and the European Research Council (ERC DLV-677555) to LB. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.