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

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

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


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.

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