Pancreas++: Automated Quantification of Pancreatic Islet Cells in Microscopy Images

Hongyu Chen,Stuart Maudsley,Bronwen Martin,Jennifer L Fiori,Josephine M Egan,Huan Cai,Sana Siddiqui

doi:10.3389/fphys.2012.00482

Abstract

The microscopic image analysis of pancreatic Islet of Langerhans morphology is crucial for the investigation of diabetes and metabolic diseases. Besides the general size of the islet, the percentage and relative position of glucagon-containing alpha-, and insulin-containing beta-cells is also important for pathophysiological analyses, especially in rodents. Hence, the ability to identify, quantify and spatially locate peripheral, and “involuted” alpha-cells in the islet core is an important analytical goal. There is a dearth of software available for the automated and sophisticated positional quantification of multiple cell types in the islet core. Manual analytical methods for these analyses, while relatively accurate, can suffer from a slow throughput rate as well as user-based biases. Here we describe a newly developed pancreatic islet analytical software program, Pancreas++, which facilitates the fully automated, non-biased, and highly reproducible investigation of islet area and alpha- and beta-cell quantity as well as position within the islet for either single or large batches of fluorescent images. We demonstrate the utility and accuracy of Pancreas++ by comparing its performance to other pancreatic islet size and cell type (alpha, beta) quantification methods. Our Pancreas++ analysis was significantly faster than other methods, while still retaining low error rates and a high degree of result correlation with the manually generated reference standard.

Highlights

Recent research has demonstrated that the maintenance of coherent somatic metabolism is vital for protecting against age or disease-related central and peripheral pathophysiology (Martin et al, 2008, 2010, 2012; Cai et al, 2012; Siddiqui et al, 2012)
This system is centered upon the pancreas, a large secretory organ possessing endocrine secretory cells that release insulin into the major circulation in response to dietary glucose
The output file contains a table with the image names, total islet area, total alpha-cell area, total alpha-cell count, interior alpha-cell area, interior alpha-cell count, alpha-cell percentage, interior alpha-cell percentage, beta-cell percentage, alpha-cell to beta-cell ratio, interior alpha-cell to beta-cell ratio, and individual islet information with respect to all the aforementioned quantifications

Summary

Introduction

Recent research has demonstrated that the maintenance of coherent somatic metabolism is vital for protecting against age or disease-related central and peripheral pathophysiology (Martin et al, 2008, 2010, 2012; Cai et al, 2012; Siddiqui et al, 2012). An appreciation of how somatic energy function is altered in aging or pathophysiological states entails at some point an in-depth analysis of the insulinotropic glucose-regulatory system. This system is centered upon the pancreas, a large secretory organ possessing endocrine secretory cells that release insulin into the major circulation in response to dietary glucose. The growth, development, function, and sensitivity of these beta-cells is, in-part, managed via a local secretory interaction with glucagon-containing alpha-cells that are present in the pancreatic islets (Jain and Lammert, 2009). Islets contain several other secretory cell types that are responsible for the local and systemic release of somatostatin (delta cells), pancreatic polypeptide (PP cells), and ghrelin (epsilon cells). The majority of the pancreatic islet mass is made up of beta- (65–80%) and alpha-(15–20%) cells and these cell populations are the most consistently measured to assess the connection between islet morphology and pancreatic function

Methods

Results

Conclusion