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Abstract：
Modern morphological and structural studies are coming to a new level by incorporating the latest methods of three-dimensional electron microscopy (3D-EM). One of the key problems for the wide usage of these methods is posed by difficulties with sample preparation, since the methods work poorly with heterogeneous (consisting of tissues different in structure and in chemical composition) samples and require expensive equipment and usually much time. We have developed a simple protocol allows preparing heterogeneous biological samples suitable for 3D-EM in a laboratory that has a standard supply of equipment and reagents for electron microscopy. This protocol, combined with focused ion-beam scanning electron microscopy, makes it possible to study 3D ultrastructure of complex biological samples, e.g., whole insect heads, over their entire volume at the cellular and subcellular levels. The protocol provides new opportunities for many areas of study, including connectomics.
摘要：
通过结合三维电子显微镜（3D-EM）的最新方法，现代形态和结构研究进入了一个新的水平。这些方法广泛应用的关键问题之一是样品制备困难。因为该方法对异质（由结构和化学成分不同的组织组成）样品的效果较差，并且需要昂贵的设备和通常需要很多时间。我们已经开发了一种简单的协议，允许在实验室中制备适用于3D-EM的异质生物样品，该实验室具有标准的电子显微镜设备和试剂供应。这个协议,结合聚焦离子束扫描电子显微镜，使得研究复杂生物样品的三维超微结构成为可能，例如，整个昆虫的头,在细胞和亚细胞水平的整个体积上。该协议为许多研究领域提供了新的机会，包括连接组学。
