Masich Sergej, Ostberg Tove, Norlén Lars, Shupliakov Oleg, Daneholt Bertil
Department of Cell and Molecular Biology, Karolinska Institutet, Box 285, SE-17177 Stockholm, Sweden.
J Struct Biol. 2006 Dec;156(3):461-8. doi: 10.1016/j.jsb.2006.05.010. Epub 2006 Jun 8.
We describe a novel approach for the accurate alignment of images in electron tomography of vitreous cryo-sections. Quantum dots, suspended in organic solvents at cryo-temperatures, are applied directly onto the sections and are subsequently used as fiducial markers to align the tilt series. Data collection can be performed from different regions of the vitreous sections, even when the sections touch the grid only at a few places. We present high-resolution tomograms of some organelles in cryo-sections of human skin cells using this method. The average error in image alignment was about 1nm and the resolution was estimated to be 5-7nm. Thus, the use of section-attached quantum dots as fiducial markers in electron tomography of vitreous cryo-sections facilitates high-resolution in situ 3D imaging of organelles and macromolecular complexes in their native hydrated state.
我们描述了一种用于玻璃体冷冻切片电子断层扫描中图像精确对齐的新方法。在低温下悬浮于有机溶剂中的量子点被直接应用于切片上，随后用作基准标记来对齐倾斜序列。即使切片仅在少数地方接触网格，也可以从玻璃体切片的不同区域进行数据采集。我们使用这种方法展示了人类皮肤细胞冷冻切片中一些细胞器的高分辨率断层扫描图。图像对齐的平均误差约为1nm，分辨率估计为5 - 7nm。因此，在玻璃体冷冻切片的电子断层扫描中使用附着在切片上的量子点作为基准标记，有助于对处于天然水合状态的细胞器和大分子复合物进行高分辨率原位三维成像。
