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. 2021 Jan 5;64(3):618–629. doi: 10.1007/s00125-020-05345-8

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Analysis of beta cell arrangement with respect to the vasculature. Human islets were fixed and the predominant cell types and vasculature were examined. (a–d) Representative images demonstrating that human islets are arranged with an alpha cell (glucagon, green) mantle and beta cell (insulin, blue) core and surrounded by a laminin capsule (red). Both (a) single and (b) multi-lobular islets retain the same structure within lobes. Linescans across a whole islet lobe show a core–mantle arrangement with bright glucagon signal close to laminin and insulin signal situated in the middle. (c) Human islets stained with insulin (blue) and glucagon (green) display a heterotypic cell arrangement along the vasculature (laminin, red). (d) Zoomed-in images showing beta cells (β) stretching through alpha cells (α) to contact the vasculature. White arrows indicate beta cell contact with vasculature. Data in (a–d) are representative of 14 islets from 3 donors. (e, f) Analysis of the distance from beta cell to the vasculature indicating that most cells are within 10 μm of the vasculature. (e) A frequency histogram showing data from individual cells (n=448 cells); (f) data presented using islet averages. Data in (e, f) is representative of 3 donors; n=6 with 2 islets analysed per donor. (g) Representative images demonstrating that beta cells (blue) elongate to make contact with islet blood vessels (red); beta cell membranes were visualised using syntaxin (green). (h) A histogram showing maximum beta cell length from individual cells (n=468 cells). (i) Maximum beta cell length presented using islet averages. Data in (h–i) are representative of 3 donors; 1–2 islets analysed per donor. (j) Insulin immunostaining has a higher intensity towards the vasculature as demonstrated by linescan analysis at the vascular and avascular face of beta cells. The red dataset on the graph represents the beta cell from the image. Data are representative of 3 donors; n=83 cells analysed using 1–2 islets per donor. Data represent mean ± SEM in (f, i). Scale bars: 50 μm on whole islet images; 10 μm on zoomed-in images. Data were collected from both pancreatomised and cadaveric donor samples. ***p<0.001. Glu, glucagon; Ins, insulin; Lam, laminin; +ve, positive