Sample Preparation and Image Processing

CEVS – Controlled Environment Vitrification System

CEVS - Controlled Environment Vitrification System

 

Leica EM BAF 060 Freeze-Fracture-Replication and cryo-SEM sample preparation System

Leica EM BAF 060 Freeze-Fracture-Replication and cryo-SEM sample preparation System

After vitrification, the specimen, mounted on a liquid nitrogen-cooled stage, is fractured in the 10-5-10-6 mbar vacuum chamber, either by a double-replica device or by a cooled knife. The fracture surface is then shadowed by Pt/C; for freeze-fracture-replication, it is also backed by a carbon layer. Metal and carbon coating is performed by two adjustable electron-sputtering guns. The coated sample is removed from the chamber. For cryo-HRSEM, the sample is transferred immediately to the HR-SEM via an evacuated liquid nitrogen cooled shuttle. In the case of freeze-fracture-replication the vitrified sample is dissolves, the replica washed, dried, and then examined by TEM at room temperature.