3-DIMENSIONAL STRUCTURE OF LIPID VESICLES EMBEDDED IN VITREOUS ICE AND INVESTIGATED BY AUTOMATED ELECTRON TOMOGRAPHY
3-DIMENSIONAL STRUCTURE OF LIPID VESICLES EMBEDDED IN VITREOUS ICE AND INVESTIGATED BY AUTOMATED ELECTRON TOMOGRAPHY
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DOI：
10.1016/s0006-3495(95)80314-0
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发表时间：
1995-04-01
影响因子：
3.4
通讯作者：
BAUMEISTER, W
中科院分区：
文献类型：
--
作者：
DIERKSEN, K;TYPKE, D;BAUMEISTER, W
自动电子断层扫描被证明是一种合适的手段来可视化嵌入在玻璃化冰中的磷脂小泡的形状。使用慢扫描电荷耦合器件相机作为记录设备，在20,000倍的放大倍数下记录60个投影的数据集所需的累积电子剂量可以保持在低至15e(-)/埃(2)(或1500电子/纳米(2))。通过三维重建的体积，在二维切片上可以清楚地看到三维重建的囊泡的膜。一些边缘指示从凝胶相冻结的囊泡的多边形形状，也被清楚地识别出来。由于目前倾斜角度范围有限(+/-60度)，在三维重建中，小泡的上、下‘帽’(约占椭圆形颗粒表面的35%)保持不可见。
Automated electron tomography is shown to be a suitable means to Visualize the shape of phospholipid vesicles embedded in vitrified ice. With a slow-scan charge-coupled device camera as a recording device, the cumulative electron dose needed to record a data set of 60 projections at a magnification of 20,000x can be kept as low as 15 e(-)/Angstrom(2) (or 1500 electrons/nm(2)). The membrane of the three-dimensionally reconstructed vesicles is clearly Visible in two-dimensional sections through the three-dimensionally reconstructed volume. Some edges indicating a polygonal shape of the vesicles, frozen from the gel phase, are also clearly recognized. Because of the presently limited tilt angle range (+/-60 degrees), the upper and lower ''caps'' of the vesicles (representing about 35% of the surface of the ellipsoidal particles) remain invisible in the three-dimensional reconstruction.