Pancreas++: Automated quantification of pancreatic islet cells in microscopy images

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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 devel-oped 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 compar-ing 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. © 2013 Chen, Martin, Cai, Fiori, Egan, Siddiqui and Maudsley.

Figures

  • FIGURE 1 | An illustration of the workflow of the Pancreas++ algorithm for determining interior alpha-cells. Contour deformation was run at 800 iterations. The islet contour was generated with linear interpolation of contour points. Interior alpha-cells were determined by the distance between each alpha-cell centroid and the nearest point in the
  • FIGURE 2 | User interface and output features. Pancreas++ output features include (left to right): image name, total islet area, total alpha-cell area, total alpha-cell count, interior alpha-cell count, interior alpha-cell area, 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. Data can be easily
  • FIGURE 3 | Validation results for quantification of interior alpha-cells. Total size of validation set was 75. The Pearson’s correlation coefficient, p-value from Pearson’s Chi-square goodness of fit test, average absolute error, and average relative error were 0.9909, 1.7×10−7, 0.8310, and 0.0158, respectively.
  • FIGURE 4 | Application of Pancreas++ in mouse, primate, and human pancreatic islet images. (A–C) Are representative images of mouse, primate, and human pancreatic islet images respectively. (D) Total islet area of mouse pancreatic islet images (arbitrary units); (E) total alpha-cell area of mouse pancreatic islet images (arbitrary units); (F) total alpha-cell numbers of mouse pancreatic islet images; (G) alpha-cell percentage of mouse pancreatic islet images; (H) beta-cell percentage of mouse pancreatic islet images; (I) total islet area of primate pancreatic islet images (arbitrary units); (J) total alpha-cell
  • FIGURE 5 | Analysis of pancreatic islets of mice on control and high fat high glucose (HFG) diet using Pancreas++, MATLAB, and manual method. (A,B) Are representative images of pancreatic islet from mice on control and HFG diets respectively. (C) Shows the time that Pancreas++, MATLAB, and manual method take to analyze the same images. The mice on HFG diet had significantly higher total islet area (D), total alpha-cell area (E), total alpha-cell numbers (F), but similar alpha-cell percentage (G) and beta-cell percentage (H) compared to the mice on control diet analyzed by
  • FIGURE 6 | Analysis of pancreatic islets from normal and diabetic primates using Pancreas++, MATLAB and manual method. (A,B) Are representative images of pancreatic islets from normal and diabetic primates respectively. (C) Shows the time that Pancreas++, MATLAB, and manual method take to analyze the same images. The diabetic primate had significantly higher total alpha-cell area (E); total alpha-cell numbers (F); alpha-cell percentage (G), lower beta-cell percentage (H), but similar islet area (D) compared to the normal primate analyzed by Pancreas++.

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APA

Chen, H., Martin, B., Cai, H., Fiori, J. L., Egan, J. M., Siddiqui, S., & Maudsley, S. (2013). Pancreas++: Automated quantification of pancreatic islet cells in microscopy images. Frontiers in Physiology, 3 JAN. https://doi.org/10.3389/fphys.2012.00482

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