title: Serial block-face imaging and its potential for reconstructing diminutive cell systems: a case study from arthropods.
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.
authors.affiliation: 1Allgemeine und Systematische Zoologie,Zoologisches Institut und Museum,Ernst-Moritz-Arndt-Universität,J.-S.-Bach-Str. 11/12,D-17487 Greifswald,Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Lipke
authors.affiliation: 2Johann-Friedrich-Blumenbach-Institute of Zoology and Anthropology,Department of Morphology,Systematics and Evolutionary Biology,Georg-August-University,Göttingen,Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hörnschemeyer
authors.affiliation: 3Gatan Inc.,Pleasanton,CA,USA.
authors.forename: Anahita
authors.initials: A
authors.lastname: Pakzad
authors.affiliation: 3Gatan Inc.,Pleasanton,CA,USA.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Booth
authors.affiliation: 1Allgemeine und Systematische Zoologie,Zoologisches Institut und Museum,Ernst-Moritz-Arndt-Universität,J.-S.-Bach-Str. 11/12,D-17487 Greifswald,Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Michalik
mesh_terms: D000818:Animals; D001181:Arthropods / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008297:Male; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D013094:Spermatozoa / Q000648:ultrastructure*