The cell in absence of aggregation artifacts.

Details

Serval ID
serval:BIB_38F45B637523
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
The cell in absence of aggregation artifacts.
Journal
Micron
Author(s)
Dubochet J., Sartori Blanc N.
ISSN
0968-4328 (Print)
ISSN-L
0968-4328
Publication state
Published
Issued date
2001
Volume
32
Number
1
Pages
91-99
Language
english
Notes
Publication types: Comparative Study ; Journal Article
Publication Status: ppublish
Abstract
Eduard Kellenberger understood that the conventional resin-embedding, he helped to develop (Ryter, A., Kellenberger, E., 1958. L'inclusion au polyester pour l'ultramicrotomie. J. Ultrastruct. Res. , 2, 200-214), was prone to aggregation artifacts (Kellenberger, E., 1987. The response to biological macromolecules and supramolecular structures to the physics of specimen cryo-preparation. In: Steinbrecht, R.A., Zierold, K. (Eds.), Cryo-techniques in Biological Electron Microscopy, Springer, Berlin, pp. 35-63). He was instrumental in developing various methods to overcome this limitation, for instance, by using low temperature-embedding and partially hydrophilic resins (Carlemalm, E., Garavito, R.M., Villiger, W., 1982. Resin development for electron microscopy and an analysis of embedding at low temperature. J. Microstruct., 126, 123-143; Villiger,W., 1993. Low temperature-embedding with Lowicryl resins. In: Robards, A.W., Wilson, A.J. (Eds.), Procedures in electron microscopy, Wiley, Chichester, UK, pp. 16:7.3-16:7.6). In principle, cryo-electron microscopy of vitreous sections is free of any aggregation artifact since the material remains fully hydrated and is free of chemical fixation or staining. The method is technically difficult still, but recent progress has made it amenable to routine practical applications. We compare here electron microscopical aspects of Zea mays meristem cells prepared by: (1) conventional resin-embedding and sectioning; (2) low temperature-embedding and sectioning of freeze substituted samples; and (3) cryo-sections of vitrified samples. The appearance of the extra-cellular space, the cytoplasm and the nucleoplasm are very different in conditions (1) and (3). They appear as compact, irregular and well delineated structures in conventional resin sections, whereas they are more diffuse and homogeneous in the vitreous sections. In the resin sections, the material seems to form a complex matrix, whereas it looks more like a thick soup in the vitreous sample. Low temperature-embedding (condition 2) shows an intermediate appearance. We suggest that regardless of the difference due to staining and different sectioning conditions, the other image differences are the consequence of aggregation artifacts in the resin-embedded specimens.
Keywords
Artifacts, Cryoelectron Microscopy, Freeze Substitution, Frozen Sections, Meristem/ultrastructure, Organelles/ultrastructure, Plastic Embedding, Zea mays/ultrastructure
Pubmed
Web of science
Create date
24/01/2008 11:25
Last modification date
20/08/2019 14:28
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