title: The cutting of ultrathin sections with the thickness less than 20 nm from biological specimens embedded in resin blocks.
abstract: Low voltage electron microscopes working in transmission mode, like LVEM5 (Delong Instruments, Czech Republic) working at accelerating voltage 5 kV or scanning electron microscope working in transmission mode with accelerating voltage below 1 kV, require ultrathin sections with the thickness below 20 nm. Decreasing of the primary electron energy leads to enhancement of image contrast, which is especially useful in the case of biological samples composed of elements with low atomic numbers. As a result treatments with heavy metals, like post-fixation with osmium tetroxide or ultrathin section staining, can by omitted. The disadvantage is reduced penetration ability of incident electrons influencing the usable thickness of the specimen resulting in the need of ultrathin sections of under 20 nm thickness. In this study we want to answer basic questions concerning the cutting of extremely ultrathin sections: Is it possible routinely and reproducibly to cut extremely thin sections of biological specimens embedded in commonly used resins with contemporary ultramicrotome techniques and under what conditions? Microsc. Res. Tech. 79:512-517, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, 37005, Czech Republic.|Faculty of Science, Charles University in Prague, Viničná 7, 128 08 Praha 2, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Nebesářová
authors.affiliation: Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Videňská, 1083, Prague, 14220, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Hozák
authors.affiliation: Institute of Scientific Instruments Academy of Sciences of the Czech Republic, Královopolská 147, Brno, 612 64, Czech Republic.
authors.forename: Luděk
authors.initials: L
authors.lastname: Frank
authors.affiliation: Delong Instruments, a.s, Palackého třída 3019, Brno, 612 00, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Štěpan
authors.affiliation: Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Branišovská 31, České Budějovice, 37005, Czech Republic.|Faculty of Sciences, University of South Bohemia at České Budějovice, Branišovská 31, České Budějovice, 37005, Czech Republic.
authors.forename: Marie
authors.initials: M
authors.lastname: Vancová
mesh_terms: D000818:Animals; D004853:Epoxy Resins / Q000737:chemistry; D004867:Equipment Design; D006321:Heart / Q000000981:diagnostic imaging; D051379:Mice; D008854:Microscopy, Electron / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy / Q000379:methods*; D009206:Myocardium / Q000648:ultrastructure; D016613:Plastic Embedding / Q000379:methods*; D011108:Polymers / Q000737:chemistry
chemical_list: D004853:Epoxy Resins; D011108:Polymers; C048709:spurr resin; C025222:Polybed 812
keywords: low voltage electron microscopy; resin embedding; ultramicrotomy; ultrathin sectioning