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Serial Block-Face Imaging and its Potential for Reconstructing Diminutive Cell Systems: A Case Study from Arthropods

Published online by Cambridge University Press:  20 February 2014

Elisabeth Lipke ,
Thomas Hörnschemeyer ,
Anahita Pakzad ,
Christopher R. Booth  and
Peter Michalik
Elisabeth Lipke*
Affiliation:
Allgemeine und Systematische Zoologie, Zoologisches Institut und Museum, Ernst-Moritz-Arndt-Universität, J.-S.-Bach-Str. 11/12, D-17487 Greifswald, Germany
Thomas Hörnschemeyer
Affiliation:
Johann-Friedrich-Blumenbach-Institute of Zoology and Anthropology, Department of Morphology, Systematics and Evolutionary Biology, Georg-August-University, Göttingen, Germany
Anahita Pakzad
Affiliation:
Gatan Inc., Pleasanton, CA, USA
Christopher R. Booth
Affiliation:
Gatan Inc., Pleasanton, CA, USA
Peter Michalik
Affiliation:
Allgemeine und Systematische Zoologie, Zoologisches Institut und Museum, Ernst-Moritz-Arndt-Universität, J.-S.-Bach-Str. 11/12, D-17487 Greifswald, Germany Research Associate, Division of Invertebrate Zoology, American Museum of Natural History, New York, USA
*
*Corresponding author. [email protected]
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Abstract

Until recently, three-dimensional reconstruction on an ultrastructural level was only possible using serial section transmission electron microscopy (ssTEM). However, ssTEM is highly challenging and prone to artifacts as, e.g., section loss and image distortions. New methods, such as serial block-face scanning electron microscopy (SBFSEM) overcome these limitations and promise a high lateral resolution. However, little is known about the usability of SBFSEM in diminutive, but highly complex cellular systems. We used spider sperm (~3 µm in diameter), which fulfills these conditions, to analyze the potential of SBFSEM compared with ssTEM. Our data suggest that the resolution obtained by SBFSEM allows depicting structures on a cellular level and is sufficient to discriminate subcellular components, but is highly dependent on previous staining procedures and electron density of the target structures.

Keywords

SBFSEMssTEMenhanced heavy metal stainingartifactspider sperm
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 3 , June 2014 , pp. 946 - 955
Copyright
© Microscopy Society of America 2014 

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