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

Heinz Horstmann, Christoph Körber, Kurt Saetzler, Daniel Aydin, Thomas Kuner

    Research output: Contribution to journalArticle

    56 Citations (Scopus)

    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.
    LanguageEnglish
    Pagese35172
    JournalPLoS ONE
    Volume7
    Issue number4
    DOIs
    Publication statusPublished - Apr 2012

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    wafers
    scanning electron microscopy
    silicon
    cells
    transmission electron microscopy
    high resolution
    automation
    ribbons
    electrons
    recording
    pixels
    lenses
    detectors

    Cite this

    Horstmann, Heinz ; Körber, Christoph ; Saetzler, Kurt ; Aydin, Daniel ; Kuner, Thomas. / Serial Section Scanning Electron Microscopy (S(3)EM) on Silicon Wafers for Ultra-Structural Volume Imaging of Cells and Tissues. In: PLoS ONE. 2012 ; Vol. 7, No. 4. pp. e35172.
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    Serial Section Scanning Electron Microscopy (S(3)EM) on Silicon Wafers for Ultra-Structural Volume Imaging of Cells and Tissues. / Horstmann, Heinz; Körber, Christoph; Saetzler, Kurt; Aydin, Daniel; Kuner, Thomas.

    In: PLoS ONE, Vol. 7, No. 4, 04.2012, p. e35172.

    Research output: Contribution to journalArticle

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