Structural biology inside multicellular specimens using electron cryotomography

Area of Science:

• Structural Biology
• Cell Biology
• Biophysics

Background:

• Electron cryomicroscopy (cryo-EM) has revolutionized structural biology.
• Electron cryotomography (cryo-ET) enables visualization of native 3D organization in biological specimens.
• Studying multicellular specimens with cryo-ET faces significant workflow challenges.

Purpose of the Study:

• To review current in situ imaging techniques for multicellular specimens using cryo-ET.
• To enumerate the limitations associated with existing cryo-ET workflows.
• To identify future advancements needed for comprehensive in situ structural biology.

Main Methods:

• Review of current techniques in sample preparation for cryo-ET.
• Analysis of data collection strategies for multicellular specimens.
• Examination of image analysis methods in cryo-ET.

Main Results:

• Latest techniques for in situ imaging of multicellular specimens are outlined.
• Limitations across sample preparation, data collection, and image analysis are detailed.
• Success stories highlight progress in cryo-ET applications.

Conclusions:

• Significant improvements are needed across the entire cryo-ET workflow for multicellular specimens.
• Future hardware and software developments are crucial for advancing in situ structural biology.
• Enhanced cryo-ET capabilities will drive new discoveries in molecular and cell biology.