System and method for preparing cryo-em grids

A method for preparing cryogenic electron microscopy samples comprising the steps of:

• A. providing a sample preparation device having:
• a cryogenically-cooled stage and a sample dispenser;
• B. defining a user-defined ice thickness;
• and
• C. depositing a liquid sample onto the stage using the sample dispenser to form a vitrified sample having a vitreous ice layer of said user-defined ice thickness.

An electron microscope (EM) preparation and imaging system comprising:

• an EM device;
• and
• a sample preparation apparatus for forming a vitreous ice layer containing a liquid sample through vitrification, the apparatus comprising:
• a cryogenically-cooled stage configured to removably receive a sample deposit surface such that the deposit surface is cryogenically cooled through direct contact with the stage;
• a sample dispenser that is at least one of laterally, longitudinally, vertically, or rotationally movable with respect to the stage and that is configured to deposit a liquid sample onto the sample deposit surface at a selected rate of deposition, wherein, once the liquid sample is deposited onto the sample deposit surface by the sample dispenser, it is vitrified automatically in place;
• and
• a sealable environment configured to be placed under at least one of a positive pressure or a negative pressure, wherein at least a portion of the EM device and sample preparation apparatus is located inside the sealable environment such that a sample may be vitrified by the sample preparation device and imaged by the EM device inside of and without being removed from the sealable environment and without changing the pressure of sealable environment.