Quick Freeze Substitution Processing of Biological Samples for Serial Block-face Scanning Electron Microscopy
Quick Freeze Substitution Processing of Biological Samples for Serial Block-face Scanning Electron Microscopy
复制标题
串行块面扫描电子显微镜生物样品的快速冷冻替代处理
DOI：
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发表时间：
2015
影响因子：
2.8
通讯作者：
Robyn Webb
中科院分区：
文献类型：
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作者：
R. Webb;Robyn Webb
直到最近，获得大面积生物样品的三维电子显微镜数据的唯一方法是通过连续切片，这是一种繁琐且技术上具有挑战性的技术。连续块面扫描电子显微镜（SBF-SEM）最近已被用作一种技术，以获得类似的结果，连续切片透射电子显微镜，但在一个更简单和半自动化的方式。它涉及到与用于透射电子显微镜类似的树脂包埋样品的工作。然而，成像是在试块表面上进行的，而不是从试块上切下的切片。图像收集是使用扫描电子显微镜中的背散射电子检测器完成的。拍摄块面的图像，然后使用金刚石刀从块上切下薄的部分。然后对新曝光的块面进行成像，从其切割另一个部分，再次成像等。成像和切片的过程反复进行，以建立三维数据集。有两种商业仪器可用于执行SBF-SEM：Gatan 3View和FEI Teneo Volumescope。
Until recently the only way to obtain three-dimensional electron microscopy data over a large area of a biological sample was via serial sectioning, a tedious and technically challenging technique. Serial block-face scanning electron microscopy (SBF-SEM) has recently been used as a technique to obtain similar results to serial sectioning transmission electron microscopy but in a simpler and semi-automated way. It involves working with a resin embedded sample similar to that used for transmission electron microscopy. However, the imaging is performed on the block-face rather than the section cut from it. Image collection is done using a backscattered electron detector in a scanning electron microscope. An image is taken of the block-face and then a thin section is cut from the block using a diamond knife. The newly exposed block-face is then imaged, another section cut from it, it is imaged again etc. This process of imaging and sectioning is performed over and over building up a three-dimensional data set. Two commercial instruments are available that will perform SBF-SEM: the Gatan 3View and the FEI Teneo Volumescope.