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Microsc Microanal. 2014 Jun;20(3):946-55. doi: 10.1017/S1431927614000087. Epub 2014 Feb 20.

Serial block-face imaging and its potential for reconstructing diminutive cell systems: a case study from arthropods.

Author information

1
1Allgemeine und Systematische Zoologie,Zoologisches Institut und Museum,Ernst-Moritz-Arndt-Universität,J.-S.-Bach-Str. 11/12,D-17487 Greifswald,Germany.
2
2Johann-Friedrich-Blumenbach-Institute of Zoology and Anthropology,Department of Morphology,Systematics and Evolutionary Biology,Georg-August-University,Göttingen,Germany.
3
3Gatan Inc.,Pleasanton,CA,USA.

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.

PMID:
24555994
DOI:
10.1017/S1431927614000087
[Indexed for MEDLINE]

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