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Updated: May 17, 2026

Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition
Published on: December 22, 2011
High-contrast observation of unstained proteins and viruses by scanning electron microscopy
1Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Central 2, Umezono, Tsukuba, Ibaraki, Japan. t-ogura@aist.go.jp
Abstract:
Scanning electron microscopy (SEM) is an important tool for the nanometre-scale analysis of the various samples. Imaging of biological specimens can be difficult for two reasons: (1) Samples must often be left unstained to observe detail of the biological structures; however, lack of staining significantly decreases image contrast. (2) Samples are prone to serious radiation damage from electron beam. Herein we report a novel method for sample preparation involving placement on a new metal-coated insulator film. This method enables obtaining high-contrast images from unstained proteins and viruses by scanning electron microscopy with minimal electron radiation damage. These images are similar to those obtained by transmission electron microscopy. In addition, the method can be easily used to observe specimens of proteins, viruses and other organic samples by using SEM.
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