Improving Cryo-Electron Tomography Data Quality and Throughput by Streamlining the Workflow

Session

Poster Session One

Authors

Marit Smeets (1), Katherine Lau (1)

Affiliations

1. Delmic B.V.

Keywords

cryo-CLEM, cryo-FIB, cryo-ET, cryo-EM

Abstract text

Cryo-electron tomography (cryo-ET) is an extremely powerful technique that allows studies of the complex structures at high resolution in a near-native state. In recent years the technique has gained popularity and an increasing number of research groups are implementing it in their lab. 

The power of cryo-ET is undisputed but the fact that the workflow is very error-prone has limited the data output¹. In the workflow, there are two main challenges: keeping the sample ice contamination-free and targeting the region of interest (ROI). 

At Delmic we developed two workflow solutions called CERES and METEOR to streamline the entire cryo-ET preparation workflow. The CERES Ice Defence System is based on the tools developed by Tacke et al.² and consists of multiple innovative tools that are tailored to minimize ice contamination during sample handling (Clean Station), transfer (Vitri-Lock) and FIB milling (Ice Shield). METEOR is an integrated fluorescence light microscope (FLM) that greatly enhances the ROI targeting inside the cryo-FIB/SEM while reducing the number of handling steps required³. 

We show that combining these innovative tools can lead to improved cryo-ET data quality and throughput. 

Figure 1: Renders of the CERES and METEOR systems: a) the optical module of METEOR b) the CERES Ice Shield protecting the sample during milling c) the CERES Clean Station d) the CERES Vitri-Lock

References

[1] Lau, K. et al. Microscopy Today 30 (2022), p. 30–35.

[2] Tacke, S. et al. Journal of Structural Biology 213 (2021), p.107743.

[3] Smeets, M. et al. Microscopy Today 29 (2021) p. 20-25