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.