Lepault J
J Microsc. 1985 Oct;140(Pt 1):73-80. doi: 10.1111/j.1365-2818.1985.tb02662.x.
Vitrified Tobacco Mosaic Virus (TMV) and T4 polyhead suspensions have been studied by cryo-electron microscopy. The order of TMV particles is preserved to distances better than 0.3 nm when vitrified. In spite of the high beam sensitivity of unstained biological material, the resolution of images extends to 1.15 nm. Radial density distributions calculated from images of TMV and the helical aggregate of TMV protein (TMVP) show that the RNA is likely visualized in the virus. However, due to the imaging conditions of unstained vitrified specimens, the position of the RNA is erroneous. While TMV and TMVP do not show any departure from cylindrical symmetry, T4 polyheads are sensitive to the thickness of the ice layer in which they are embedded. To avoid flattening, T4 polyheads have to be embedded in an ice layer thicker than their diameter.
已通过冷冻电子显微镜对玻璃化烟草花叶病毒(TMV)和T4多面体悬浮液进行了研究。玻璃化时,TMV颗粒的排列在距离大于0.3 nm的情况下得以保留。尽管未染色的生物材料对电子束敏感性很高,但图像分辨率仍可达1.15 nm。从TMV图像和TMV蛋白(TMVP)的螺旋聚集体计算出的径向密度分布表明,病毒中的RNA可能被可视化。然而,由于未染色玻璃化标本的成像条件,RNA的位置存在误差。虽然TMV和TMVP没有显示出与圆柱对称性的任何偏差,但T4多面体对其所嵌入的冰层厚度很敏感。为避免扁平化,T4多面体必须嵌入比其直径更厚的冰层中。