Microfluidic Cryo-EM Grid Preparation

  • The single-cell analysis and grid preparation platform consists of multiple motorized stages and syringe pumps, assembled around an inverted microscope.

  • Individual modules are developed in LabVIEW to control the different hardware components of the analysis platform. The OpenBEB software framework is used to coordinate the independent modules for process automation.

About-+

We are developing new methods for the loss-less (no blotting steps) sample preparation of nanoliter volumes for negative stain and cryo-EM. Our work includes hardware and software development. These methods are then used to look at the proteome of single mammalian cells by electron microscopy.

More detail+-

In recent years there has been a renaissance of electron microscopy (EM) in biological research. While image acquisition and processing have largely benefited from new direct electron detection (DED) cameras, sample preparation is still in the pre-DED era. Current methods suffer from two major drawbacks: i) high sample consumption (around 3 µl/grid) and ii) massive blotting steps that remove more than 99% of the sample. In addition, these blotting steps are known to generate unrepresentative sample populations on the EM grid.

We are developing methods to reproducibly deposit a nanolitre volume of sample onto an EM grid at room and at cryo-temperatures.