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We share answers to frequently asked questions about sample preparation for cryo-EM

What grids can be used?

Basically, we can process all grids that allow pre-clipping in Autogrid carrier rings. When depositing sample, the writing pin does not touch the actual grid, so there is no physical contact between pin and grid. To date, we have not encountered damage of grids due to shear force when depositing sample.

Can we modify settings for plasma treatment?

The plasma treatment of grids is integrated to ensure reproducibility (timings, type of treatment, etc.) between grids. However, if you prefer to treat your grids in a specific way, the step inside the VitroJet can be bypassed. The VitroJet will be equipped with a plasma cleaner, allowing for the use of different gases.

How many grids can we produce in a single session?

The VitroJet is equipped with a cassettes holding up to 12 pre-clipped grids. Normally, this is the maximum number of grids used in a single session. Of course, if you place another storage cassette in the VitroJet after the first one is finished, you can process as many as you like, and operation is limited by the contamination of your cryogenic compartment.

How long does the nitrogen/ethane last?

The VitroJet is equipped with a system that automatically refills both liquid nitrogen and ethane. The degree of contamination determines how long you can work with the system. Use in a low-humidity environment is preferred.

Are we restricted to a type or brand of microscope?

The VitroJet is designed for optimal control and reproducibility, therefore pre-clipped grids are used because they allow automated handling. Although pre-clipped grids are used, (side-entry) cryoholders can be adjusted with only minor changes to fit pre-clipped grids. Contact CryoSol for further information about compatibility with your microscope or cryoholder.

Are we restricted to only one sample per session?

No, you’re not. You can change your sample at any point in the session. This applies to any other parameters the user wishes to adjust in the session.

Where can we place the VitroJet?

The VitroJet is designed as a bench-top device, it has a footprint of approximately 80 x 80 cm / 32 x 32 inch. Although the VitroJet is a closed system, ideally the environment has a relative low humidity and limited flow, to ensure minimal ice contamination.

What are the requirements with respect to connections to the VitroJet?

The VitroJet is designed to operate at 230 VAC/50 Hz and/or 110 VAC/60 Hz. A dry nitrogen gas connection is required for drying. Depending on your plasma cleaning requirements, you may want to connect other cleaning gases. Distilled water is used for obtaining the humidity in the sample deposition module.