Cryo sample preparation
Please see CAM: Biological sample preparation for more biological specimen preparation facilities. In addition to LN2 Cold stages on 2 SEMs and 3 TEMs, CAM has the following cryo preparation equipment available:
Leica EM-ICE High pressure freezer
- Pressure controllable to 2300 bar. Specimen holders currently c. 3mm or 6mm dia x 200 micron deep larger flat holders.
- High pressure freezing, instantly immobilising cell contents and preserving fine structure and antigenicity by rapid freezing under pressure.
- This is a high pressure freezing system for vitrifying samples up to 300µm in thickness without the artefacts of chemical fixation.
Reichert AFS automated freeze substitution unit
This is an automatic user-programmed substitution and low temperature embedding system for cryo-prepared samples:
- Working range: -140 to +65°C
- Time settings: 0 to 999.9 hr for each step
- 3 Holding-Time / Temperature settings
- 2 Slope settings i.e. time between two temperatures
- 10 Programme memories
Leica EM AFS2
Automated freeze substitution unit allow freeze substitution and low temperature resin polymerization.
- Automatic reagent handling system
- Stereomicroscope attached for viewing samples
Plunge Freezer
This is a plunge freezer for the preparation of vitrified thin samples for cryo-TEM.
- Adjustable plunge rod with attached tweezer.
FEI Vitrobot
A high-performance, fully automated instrument used to prepare vitrified samples for cryo-EM imaging in a reproducible, standardised manner. After applying the sample to the grid, the Vitrobot automatically blots away excess liquid, and rapidly freezes the sample in liquid ethane. The entire process is performed in a temperature- and humidity-controlled environment, which minimises sample contamination/degradation and maximises sample quality.