Achieve Precise Cryo-ET Lamella Preparation with Tescan Cryo-FIB-SEMs

Why Reliable Cryo-ET Lamella Preparation Is Essential for High-Resolution Cryo-ET

High-resolution cryo-electron tomography relies on the quality of lamellae—ultra-thin slices of frozen biological material. But lamella preparation has remained one of the most demanding steps in the workflow.

If lamellae are too thick, contaminated, or damaged during milling, data quality is compromised. With exceptional ion beam stability ensuring efficient material removal and minimal damage during final polishing, the workflow delivers consistently high-quality results. Stable cryogenic conditions are also essential to preserve delicate, frozen-hydrated structures. Tescan Cryo FIB-SEMs are developed to maintain sample integrity throughout the entire preparation process, featuring:

• Stable ion beam performance for efficient rough milling and precise low-kV polishing

• Integrated cryo-stage (Leica or Quorum) to ensure stable cryogenic conditions throughout the workflow

• METEOR 2.0, integrated fluorescence microscope for precise targeting, reduced sample handling and contamination risk, and faster overall operation

Combined, they deliver the stability and control essential for producing high-quality cryo lamellae that enable reliable cryo-ET reconstruction.

How Cryo Lamellae Are Prepared Using Tescan Cryo-FIB-SEM

Vitrified cells cultured on TEM grids were sputter-coated with platinum and transferred to the Tescan Cryo-FIB-SEM. On-grid lamella preparation was performed using the integrated Leica/Quorum cryo-stage and cryo-shields, with CORAL™ software guiding region-of-interest (ROI) selection.

A protective platinum layer was deposited using the Gas Injection System (GIS) to protect the surface and reduce artifacts. Trenches were then milled with precise, controlled material removal.

The final lamella was finely polished using low kV. Lamellae were subsequently recoated and transferred directly to the cryo-TEM or placed in storage for further analysis. This workflow enabled researchers to achieve consistent, high-quality lamellae suitable for high-resolution cryo-ET.

Consistent Lamella Quality for Reliable Cryo-ET Data

Using Tescan Cryo-FIB-SEM systems, researchers can produce high-precision, planar lamellae optimized for high-resolution cryo-ET. The combination of low-kV, low-dose milling in stable cryogenic conditions enables precise thinning and minimizes artifacts. This preserves delicate subcellular structures for downstream imaging.

Consistent lamella thickness and structural integrity are maintained across sites on the same grid, highlighting the reproducibility of the cryo FIB-SEM workflow.

This combination of precision milling, cryo-stable hardware, and targeted site selection streamlines lamella preparation while maintaining sample integrity.