Screening for two-dimensional crystals by transmission electron microscopy of negatively stained samples.

Screening for two-dimensional crystals by transmission electron microscopy of negatively stained samples.
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通过负染色样品的透射电子显微镜筛选二维晶体。

DOI:
10.1007/978-1-62703-176-9_5
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发表时间:
2013
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Schmidt-Krey,Ingeborg
Schmidt-Krey,Ingeborg
中科院分区:
--
文献类型:
--
作者:
Dreaden,TinaM;Metcalfe,Maureen;Kim,LauraY;Johnson,MatthewC;Barry,BridgetteA;Schmidt-Krey,Ingeborg

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通过电子晶体学对可溶性蛋白和膜蛋白进行结构研究包括几个关键步骤。虽然二维 (2D) 结晶可以说是电子晶体学的主要瓶颈,但通过透射电子显微镜 (EM) 筛选来识别 2D 晶体需要非常小心和实践。与第一有序阵列的识别相比,样品制备和 EM 都是相对容易获得的技能。除此之外,膜可以具有多种形态和尺寸。在这里,我们描述了二维晶体筛选以及样品评估所涉及的所有步骤。
Structural studies of soluble and membrane proteins by electron crystallography include several critical steps. While the two-dimensional (2D) crystallization arguably may be described as the major bottleneck of electron crystallography, the screening by transmission electron microscopy (EM) to identify 2D crystals requires great care as well as practice. Both sample preparation and EM are skills that are relatively easily acquired, compared to the identification of the first ordered arrays. Added to this, membranes may have a variety of morphologies and sizes. Here we describe all steps involved in the screening for 2D crystals as well as the evaluation of samples.
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