We have made a small contribution to Jan Felix’s excellent cryoEM work by optimizing grids to overcome preferred particle orientation problems and enabling high-resolution protein structure determination. Specifically, we addressed two key challenges:

1. Overcoming Preferred Orientation with Graphene Grids

Using PUXANO graphene grids, the GARP:TGF-β1:Fab complex could be stabilized, which reduced complex disassembly and already partially improved particle orientation< compared to standard holey grids.

2. Achieving Ultra-Thin Ice and Tilted Data Collection

Minimizing ice thickness during vitrification resulted in grids with ultra-thin ice (~30 nm). This enabled data collection at 0°, 16°, and 35° tilt angles, yielding maps at 2.89–3.0 Å resolution.

 

This was the first time our graphene grids were combined with tilted data collection, and what a nice result. A similar approach has now also been applied for a tomography study using subtomogram averaging (results to be published soon). You can Read the full Cell Reports article here.

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