Abstract text (incl. figure legends and references)
Recent developments in the electron microscopy (EM) field have led to a step change in our ability to solve the "high resolution" structure of previously intractable systems. This is exemplified by the sudden expansion in membrane protein structures solved revealing new insights into structure, mechanism and regulation. Advances in the size of data sets that can be collected and processed by electron microscopy (EM) and the broad range of samples that can be studied makes it ideally suited to time-resolved studies. This talk will describe the ongoing developments of a cryo-electron microscopy grid preparation device that permits rapid mixing and vitrification of samples within ~10 ms allowing us to pull out non-equilibrium states on a range of systems from membrane proteins to viruses. Moreover, rapid grid preparation has also provided new insights into factors that can be detrimental to sample preparation and the talk will discuss ways to improve grid making for single particle cryoEM. A significant focus of the talk will be in our new findings in factors that are detrimental to sample preparation, especially those caused by interactions with the air-water-interface. The talk will conclude with our current overview of the challenges in sample preparation for both conventional cryoEM and time-resolved studies and look towards future developments in the field.