Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy
Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy
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DOI：
10.3791/59480
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发表时间：
2019-08-01
影响因子：
1.2
通讯作者：
Lippens, Saskia
中科院分区：
文献类型：
--
作者：
Guerin, Christopher J.;Kremer, Anna;Lippens, Saskia
该方案允许在电子显微镜的分辨率水平上对细胞或组织样本进行高效和有效的三维成像。多年来，电子显微镜(EM)一直是一种固有的二维技术。随着连续扫描电子显微镜成像技术(体积EM)的出现，使用集成切片机或聚焦离子束对嵌入的组织进行切片并观察，三维变得容易实现。连续块面扫描电子显微镜(SBF-SEM)采用封闭在扫描电子显微镜小室内的超微切割机。它有能力处理大样本(1,000微米x 1,000微米)，并在小X，Y像素尺寸下拍摄大视场图像，但受到钻石刀在Z维度的限制。聚焦离子束扫描电子显微镜(FIB-SEM)不限于3D分辨率，(各向同性体素
This protocol allows for the efficient and effective imaging of cell or tissue samples in three dimensions at the resolution level of electron microscopy. For many years electron microscopy (EM) has remained an inherently two-dimensional technique. With the advent of serial scanning electron microscope imaging techniques (volume EM), using either an integrated microtome or focused ion beam to slice then view embedded tissues, the third dimension becomes easily accessible. Serial block face scanning electron microscopy (SBF-SEM) uses an ultramicrotome enclosed in the SEM chamber. It has the capability to handle large specimens (1,000 mu m x 1,000 mu m) and image large fields of view at small X,Y pixel size, but is limited in the Z dimension by the diamond knife. Focused ion beam SEM (FIB-SEM) is not limited in 3D resolution, (isotropic voxels of