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Serial Section Scanning Electron Microscopy (S(3)EM) on Silicon Wafers for Ultra-Structural Volume Imaging of Cells and Tissues.

Biomedical Sciences Research Institute Computer Science Research Institute Environmental Sciences Research Institute Nanotechnology & Advanced Materials Research Institute

Horstmann, Heinz, Körber, Christoph, Saetzler, Kurt, Aydin, Daniel and Kuner, Thomas (2012) Serial Section Scanning Electron Microscopy (S(3)EM) on Silicon Wafers for Ultra-Structural Volume Imaging of Cells and Tissues. PLoS One, 7 (4). e35172. [Journal article]

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URL: http://dx.doi.org/10.1371/journal.pone.0035172

DOI: 10.1371/journal.pone.0035172

Abstract

High resolution, three-dimensional (3D) representations of cellular ultrastructure are essential for structure function studies in all areas of cell biology. While limited subcellular volumes have been routinely examined using serial section transmission electron microscopy (ssTEM), complete ultrastructural reconstructions of large volumes, entire cells or even tissue are difficult to achieve using ssTEM. Here, we introduce a novel approach combining serial sectioning of tissue with scanning electron microscopy (SEM) using a conductive silicon wafer as a support. Ribbons containing hundreds of 35 nm thick sections can be generated and imaged on the wafer at a lateral pixel resolution of 3.7 nm by recording the backscattered electrons with the in-lens detector of the SEM. The resulting electron micrographs are qualitatively comparable to those obtained by conventional TEM. S(3)EM images of the same region of interest in consecutive sections can be used for 3D reconstructions of large structures. We demonstrate the potential of this approach by reconstructing a 31.7 µm(3) volume of a calyx of Held presynaptic terminal. The approach introduced here, Serial Section SEM (S(3)EM), for the first time provides the possibility to obtain 3D ultrastructure of large volumes with high resolution and to selectively and repetitively home in on structures of interest. S(3)EM accelerates process duration, is amenable to full automation and can be implemented with standard instrumentation.

Item Type:Journal article
Faculties and Schools:Faculty of Life and Health Sciences
Faculty of Life and Health Sciences > School of Biomedical Sciences
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Molecular Medicine
ID Code:22071
Deposited By:Dr Kurt Saetzler
Deposited On:15 May 2012 13:01
Last Modified:15 May 2012 13:01

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