The cutting of ultrathin sections with the thickness less than 20 nm from biological specimens embedded in resin blocks |
| |
Authors: | Jana Nebesá?ová Pavel Hozák Luděk Frank Petr ?těpan Marie Vancová |
| |
Affiliation: | 1. Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, ?eské Budějovice, Czech Republic;2. Faculty of Science, Charles University in Prague, Czech Republic;3. Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic;4. Institute of Scientific Instruments Academy of Sciences of the Czech Republic, Brno, Czech Republic;5. Delong Instruments, Brno, Czech Republic;6. Faculty of Sciences, University of South Bohemia at ?eské Budějovice, ?eské Budějovice, Czech Republic |
| |
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. |
| |
Keywords: | low voltage electron microscopy resin embedding ultramicrotomy ultrathin sectioning |
|
|