Difference: CemProtocEC (12 vs. 13)

Revision 1326 Oct 2009 - Main.KdDerr

 
META TOPICPARENT name="CemProtoc"
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Principles & Protocols

Principles - Electron crystallography

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In order to visualize a sample in a transmission electron microscope the sample must (a) be thin enough such that a beam of electrons can penetrate it (<250nm, ideally <100nm), (b) be deposited onto an EM grid, which is a thin circular copper grid that is 3mm in diameter, and (c)withstand high vacuum and electron radiation within the microscope column. For tissue, samples are prepared by cutting thin sections

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<		 (sectioning). For aqueous suspensions of macromolecules, including
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>		 (sectioning). For aqueous suspensions of macromolecules, including
  2D crystals and ordered helical arrays, 1-5 microliters of the solution is pipetted onto the EM grid, which is then subjected to either
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<		 negative staining, plunge freezing, or a combination of these sample preservation techniques,
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>		 negative staining, plunge freezing, or a combination of these sample preservation techniques, cryo-negative staining.
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<		 cryo-negative staining.
 Two dimensional crystals are commonly formed by membrane proteins, such as bacteriorhodopsin or aquaporins. These crystals ideally consist of a single layer of molecules that are held together by specific contacts, showing crystalling order.

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-- KakoliMitra - 11 Feb 2009

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