Enabling a full Cryo-ET workflow with the TESCAN S8000G FIB-SEM

Jun/11/2018
Cryo FIB-SEM: A leap into the ultrastructure of biological specimens
Cryo-electron tomography (Cryo-ET) is an established method with increasing popularity to study molecules, macromolecular complexes and cellular ultrastructure. Focused Ion Beam (FIB) has been used in recent years to overcome the limitations of conventional cryo-sectioning and sample preparation for Cryo-ET. FIB enables the preparation of artefact-free thin lamellae of vitrified biological specimens, ideal for subsequent successful TEM analysis and tomography. The TESCAN cryo-workflow consisting of the TESCAN S8000G FIB-SEM system equipped with a Leica cryo-preparation equipment is a concrete solution for in-situ preparation of site-specific thin lamella from a fully hydrated vitrified sample.

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Application Example

Cryo FIB-SEM-A leap into the ultrastructure of biological specimens
 
Cryo-electron tomography (Cryo-ET) is an established method with increasing popularity to study molecules, macromolecular complexes and cellular ultrastructure. The principal limitation of the technique is the quality and thickness of the specimen. Introducing artefacts during the cryo sample preparation strongly influences the obtained results. To overcome the disadvantages of cryo-ultramicrotomy, a cryo FIB-SEM is used as a powerful tool for preparation of fine cryo-TEM samples. Using cryo FIB-SEM as a part of the whole cryo-TEM workflow, subcellular structure of the cells can be revealed in a form of a planar lamella (on-grid lamella). The cryo on-grid lamella is prepared without cutting artefacts and is thin enough for transmitted electrons. One of the key advantages of the cryo-FIB is the ability to prepare site-specific lamellas based on correlation with the data from light microscopic methods. By bringing the functional and ultrastructural data together dramatically helps us better understand the complexity of the biological systems.