Evaporation during preparation of unsupported thin vitrified aqueous layers for cryo-electron microscopy
- PMID: 2250188
- DOI: 10.1002/jemt.1060160407
Evaporation during preparation of unsupported thin vitrified aqueous layers for cryo-electron microscopy
Abstract
Evaporation of water cannot be fully avoided when an unsupported thin vitrified film of an aqueous suspension is prepared for cryo-electron microscopy. This results in increasing concentration of solute which could affect the observed material. We have quantitatively studied this effect by measuring the contrast of polystyrene spheres in a metrizamide solution. The drying effect is generally negligible when specimens are prepared on a hydrophilic perforated support but it is frequently important when hydrophobic films are used instead. A flow of humid air, double blotting with minimal exposure of the thin liquid film to the atmosphere, or an automatic plunger optimizing the blotting conditions are simple methods for reducing drying effects. With this third device acting on a hydrophilic supporting film, the increase of solute concentration is limited to less than 20%.
Similar articles
-
The ultrastructure of cryo-sections and intact vitrified cells--the effects of cryoprotectants and acceleration voltage on beam induced bubbling.Scanning Microsc Suppl. 1991;5(4):S43-51; discussion S51-2. Scanning Microsc Suppl. 1991. PMID: 1822023
-
Thin films composed of multiwalled carbon nanotubes, gold nanoparticles and myoglobin for humidity detection at room temperature.Chemphyschem. 2007 Feb 2;8(2):264-9. doi: 10.1002/cphc.200600409. Chemphyschem. 2007. PMID: 17221901
-
DNA-DNA interaction in thin layer analysed by cryo-electron microscopy.C R Acad Sci III. 1995 Oct;318(10):1015-20. C R Acad Sci III. 1995. PMID: 8556446
-
Self-assembling structures and thin-film microscopic morphologies of amphiphilic rod-coil block oligomers.J Colloid Interface Sci. 2005 Sep 15;289(2):488-97. doi: 10.1016/j.jcis.2005.03.052. J Colloid Interface Sci. 2005. PMID: 15936765
-
Cryo-electron microscopy of vitreous sections.EMBO J. 2004 Sep 15;23(18):3583-8. doi: 10.1038/sj.emboj.7600366. Epub 2004 Aug 19. EMBO J. 2004. PMID: 15318169 Free PMC article. Review.
Cited by
-
Cryo-electron microscopy and three-dimensional reconstruction of the calcium release channel/ryanodine receptor from skeletal muscle.J Cell Biol. 1994 Oct;127(2):411-23. doi: 10.1083/jcb.127.2.411. J Cell Biol. 1994. PMID: 7929585 Free PMC article.
-
Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles.Nucleic Acids Res. 2009 May;37(9):2882-93. doi: 10.1093/nar/gkp137. Epub 2009 Mar 12. Nucleic Acids Res. 2009. PMID: 19282451 Free PMC article.
-
Cooperative kinking at distant sites in mechanically stressed DNA.Nucleic Acids Res. 2011 Dec;39(22):9820-32. doi: 10.1093/nar/gkr666. Epub 2011 Sep 14. Nucleic Acids Res. 2011. PMID: 21917856 Free PMC article.
-
Localization of calmodulin binding sites on the ryanodine receptor from skeletal muscle by electron microscopy.Biophys J. 1994 Dec;67(6):2286-95. doi: 10.1016/S0006-3495(94)80714-3. Biophys J. 1994. PMID: 7696469 Free PMC article.
-
A novel immunogold cryoelectron microscopic approach to investigate the structure of the intracellular and extracellular forms of vaccinia virus.EMBO J. 1996 May 15;15(10):2343-55. EMBO J. 1996. PMID: 8665841 Free PMC article.