A novel 2D and 3D method for automated insulin granule measurement and its application in assessing accepted preparation methods for electron microscopy

Electron Microscopy and Analysis Group Conference 2013

Mantell, J.^1 ; Nam, D.^2 ; Bull, D.^2 ; Achim, A.^2 ; Verkade, P.^1,3

Wolfson Bioimaging Facility, University of Bristol, Medical Sciences, University Walk, Bristol BS8 1TD, United Kingdom^1

Visual Information Laboratory, University of Bristol, Merchant Venturers Building, Woodland Road, Bristol BS8 1UB, United Kingdom^2

Schools of Biochemistry and Physiology and Pharmacology, University of Bristol Medical Sciences, University Walk, Bristol BS8 1TD, United Kingdom^3

关键词: 2-D and 3-D methods;    High-pressure freezing;    Islets of Langerhans;    Manual measurements;    Preparation method;    Sample preparation;    Tomographic reconstruction;    Transmission electron microscopy images;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/522/1/012022/pdf
DOI  :  10.1088/1742-6596/522/1/012022

来源: IOP

【 摘 要 】

Transmission electron microscopy images of insulin-producing beta cells in the islets of Langerhans contain many complex structures, making it difficult to accurately segment insulin granules. Furthermore the appearance of the granules and surrounding halo and limiting membrane can vary enormously depending on the methods used for sample preparation. An automated method has been developed using active contours to segment the insulin core initially and then expand to segment the halos [1]. The method has been validated against manual measurements and also yields higher accuracy than other automated methods [2]. It has then been extended to three dimensions to analyse a tomographic reconstruction from a thick section of the same material. The final step has been to produce a GUI and use the automated process to compare a number of different electron microscopy preparation protocols including chemical fixation (where many of halos are often distended) and to explore the many subtleties of high pressure freezing (where the halos are often minimal, [3]).