Low-vibration and drift miniature cryogenic stage for Cryo-EM applications.

Pieter-Paul Lerou

Demcon kryoz, Enschede, The Netherlands

pieter.lerou@demcon.nl

The field of cryo-electron microscopy (cryo-EM) has seen a large growth the past decade. With the goal of workflow optimization, the cryo-EM community has an urge for new approaches in sample preparation. One of the complicated steps in the cryo TEM workflow is the thinning down of the cryogenically cooled sample (such as cells, bacteria and tissues), using a FIB/SEM (a focused ion beam combined with a scanning electron microscope), before yielding results in the cryo TEM. To pinpoint the region of interest, a fluorescence microscope is used during the FIB/SEM process. The setup requires a cryocooler to keep the sample below a certain temperature where the ice remains in its amorphous state, which is crucial for the process to succeed. We present the development of a miniature cryogenic stage that can cool these type of samples during the FIB/SEM/FM process with the challenging requirements of ultra-low thermal drift and mechanical vibrations.