Difference: CemProtocEC (11 vs. 12)
Revision 1213 Oct 2009 - Main.KdDerr
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| > > | Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM | |||||||
Principles & Protocols | ||||||||
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| > > | Principles - Electron crystallographyJump to Protocol | |||||||
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| < < | Electron crystallography | |||||||
| 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
(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
negative staining,
plunge freezing, or
a combination of these sample preservation techniques,
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. Public website mirror
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| History: r14 < r13 < r12 < r11
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