CryoGO enables high-resolution structural profiling of endogenous cellular macromolecules.

Area of Science:

• Structural Biology
• Cell Biology
• Cryo-electron Microscopy

Background:

• Understanding cellular function requires visualizing macromolecular structures in their native environment.
• Current methods for preparing cellular specimens for high-resolution imaging are limited in accessibility and throughput.
• Bridging the gap between molecular architecture and physiological function remains a challenge.

Purpose of the Study:

• To introduce a novel, rapid, and scalable method for preparing cell-derived specimens for cryo-electron microscopy.
• To enable high-resolution structure determination of endogenous macromolecular assemblies within their native cellular context.
• To facilitate time-resolved structural profiling of cellular processes.

Main Methods:

• cryoGO (on-Grid Opening cryo-electron microscopy) mechanically disrupts cells directly on EM grids.
• The method produces cell-derived specimens suitable for high-resolution single-particle cryo-EM.
• Compatible with standard cryo-EM infrastructure and requires minimal cell numbers.

Main Results:

• Achieved near-atomic resolution structure determination of diverse endogenous macromolecular assemblies.
• Captured the conformational and compositional landscape of ribosomes, preserving cellular state distributions and spatial heterogeneity.
• Demonstrated the capability for time-resolved structural profiling on timescales of seconds.

Conclusions:

• cryoGO significantly lowers the technical barrier to high-resolution native structural biology.
• The method provides unprecedented insights into the structural dynamics and heterogeneity of cellular components.
• Enables rapid structural profiling for studying cellular adaptation and remodeling.