Vitrobot Mark IV - Automated Vitrifier | NTU Institute of Structural Biology

The Vitribot Mark IV is used for CryoEM sample preparation. It enables users to reproducibly vitrify biological samples for cryoelectron microscopy.

If you are interested in using the equipment or require assistance and training, please kindly contact Dr. Boo Zhao Zhi.

Sample application

Small sample volumes can be applied manually with a pipette through a small side port on the left and the right side of the climate chamber
Both application time and wait time (between application and blotting) are software controlled and can be set in the user interface
Precisely timed control of multiple sample applications, blotting actions, and vitrification enables time resolved analysis of interactions among separately applied components"

Blotting device

Excess fluid is removed from the grid by (repeated) blotting with filter paper on rotating foam pads
Number of blotting actions (max. 16 times for one grid) and duration of blotting are software controlled and can be set in the user interface
Longitudinal grid positioning (‘blot offset’) and wait time between blotting and vitrification (‘drain time’) are user definable

Vitrification process

Automated shutter control allows smooth, instant injection of the sample grid into the coolant (liquid ethane or propane). A lift for the container brings the coolant close to the shutter to ensure optimal vitrification
Synchronous lowering of the coolant container and the grid holder keeps the grid submerged in the coolant and minimizes the risk of contamination prior to transfer ring the sample into a storage box or cryo holder
Coolant container with integrated anti-contamination ring z
Grid transfer from the coolant towards a grid box in the liquid nitrogen environment is semi automated

• Cryo-Plunger

• Working temperature 4-60 °C

• Peltier controlled heating/cooling

• Maintains relative humidity at 100%

• Ultrasonic controlled humidification

Vitrobot Mark IV - Automated Vitrifier