MICROSCOPY Vol.47▶No.1 2012
â– Lectures

Cryo-Electron Microscopy of Vitreous Sections

Kenji Iwasaki, Naoyuki Miyazaki, Yoshiko Ito and Junichi Takagi

Abstract: Cryo-electron microscopy of vitreous sections (CEMOVIS) enables us to observe hydrated biological materials that are cut into thin sections following physical fixation by high-pressure freezing. Thin sectioning conventionally embedded specimens causes certain artifacts, including a lack of molecules that are unfixed and deformation of molecules through chemical fixation, dehydration, and heavy metal staining. On the other hand, by avoiding the abovementioned processes, vitrification yields specimens that are closer to the native state. However, some artifacts caused by direct sectioning of vitrified ice have been verified since this technique first appeared. Here, we introduce the pros and cons of CEMOVIS, the methods for utilization of CEMOVIS, and the protocol and materials necessary to implement it.

Key words: CEMOVIS, Cryo-electron microscopy, high-pressure freezing, vitreous