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Vitreous cryo‐sectioning of cells facilitated by a micromanipulator
Author(s) -
LADINSKY MARK S.,
PIERSON JASON M.,
McINTOSH J. RICHARD
Publication year - 2006
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2006.01674.x
Subject(s) - micromanipulator , microtome , ribbon , materials science , microscopy , optical sectioning , electron microscope , cryo electron microscopy , transmission electron microscopy , optics , chemistry , composite material , nanotechnology , computer science , biochemistry , physics , artificial intelligence
Summary Sectioning vitrified cells and tissues for cryo‐electron microscopy is more challenging than room‐temperature sectioning of plastic‐embedded samples. As the sample must be kept very cold (<−130 °C) and because there is no liquid upon which the sections can float as they are cut, transferring the sections from the knife edge to a grid is one of the more difficult steps in the process. We employed a micromanipulator to hold and control the cryo‐sections as they come off the knife. This allows slower cutting speeds than are typically used in vitreous cryo‐sectioning and contributes to better control during cutting, which facilitates repeatable placement of a ribbon of sections onto a grid. The ribbon is kept under tension during the entire cutting process, which may decrease folding and/or compression, features that are inherent to vitreous sections. Furthermore, the added control afforded by this technique makes it easier for multiple ribbons to be placed on a single grid, thereby increasing the number of sections that can be examined and imaged during a microscopy session. It even allows for serial cryo‐electron microscopy. As such, this approach is an advance in the cryo‐microtomy of vitreous sections.