Cryo-electron microscopy of vitreous sections.

Area of Science:

• Structural molecular biology
• Cell biology
• Biophysics

Background:

• Cryo-electron microscopy (cryo-EM) of thin films enables native state visualization of biological particles since the 1980s.
• 3-D reconstruction combined with cryo-EM is a key tool in structural molecular biology.
• Observing larger biological structures like cells and tissues in their native state presents challenges due to film thickness limitations.

Purpose of the Study:

• To introduce and detail the methodology of Cryo-electron microscopy of vitreous sections (CEMOVIS).
• To highlight the advantages of CEMOVIS for observing biological samples in their native state.
• To discuss the potential of CEMOVIS, especially when combined with electron tomography.

Main Methods:

• Vitrification of sizable biological material.
• Sectioning vitrified samples into ultrathin sections.
• Observation of ultrathin sections in the vitrified state using cryo-electron microscopy.

Main Results:

• CEMOVIS overcomes the limitations of thin-film cryo-EM for larger biological specimens.
• The native state of biological samples is visualized with greater detail than previously possible.
• Technical difficulties associated with vitrification and sectioning have been resolved.

Conclusions:

• CEMOVIS provides a solution for observing cells and tissues in their native, hydrated state.
• The technique offers significantly improved detail and a more accurate representation of biological structures.
• Combining CEMOVIS with computerized electron tomography will unlock its full potential for 3-D reconstruction and structural analysis.