Preparation of Sample Support Films in Transmission Electron Microscopy using a Support Floatation Block

Area of Science:

• Structural Biology
• Biophysics
• Microscopy Techniques

Background:

• Cryo-electron microscopy (cryo-EM) is crucial for determining macromolecular structures.
• Sample preparation, particularly avoiding air-water interface denaturation, is a major bottleneck in cryo-EM.
• Support films (carbon, graphene, graphene oxide) mitigate air-water interface issues but are difficult to handle.

Purpose of the Study:

• To introduce a novel, user-friendly floatation block for cryo-EM support film preparation.
• To streamline the process of applying delicate support films to EM grids.
• To improve the quality and reproducibility of cryo-EM sample preparation.

Main Methods:

• Development and application of a floatation block for wetted transfer of support films.
• Step-by-step protocols for preparing amorphous carbon, graphene, and graphene oxide supports.
• Minimizing manual handling steps to preserve support film integrity and reduce contamination.

Main Results:

• The floatation block facilitates easy and efficient transfer of support films.
• Reduced handling preserves the physical integrity of delicate support films.
• Minimized contamination allows for optimal thin ice film generation for cryo-EM imaging.

Conclusions:

• The developed floatation block significantly simplifies cryo-EM support film preparation.
• This method enhances sample quality by reducing denaturation and improving ice formation.
• The protocols provide a reproducible approach for preparing various support films for cryo-EM.