Abstract
Cryo-Electron Microscopy (Cryo-EM) is in the process of revolutionising the study of biological molecules. However, the preparation of the sample usually takes at least a few seconds, precluding the study of short-lived (10 – 1000 millisecond) states in a reaction. We have built upon a technique introduced by Lu et al. (2009) who developed a microfluidic chip that mixes two components, allows the mixture to react for a defined time, and sprays the product onto the EM grid as the latter is propelled into the cryogen. In our hands we have obtained the first results demonstrating that we can capture short-lived states in an in vitro bacterial translation system during translation initiation, termination, and recycling.
Suggested reading:
Frank, J. (2017). Time-resolved cryo-electron microscopy: Recent progress. J. Struct. Biol. 200, 303-306.
Lu, Z. et al. (2009). Monolithic microfluidic mixing-spraying devices for time-resolved cryo-electron microscopy. J. Struct. Biol. 168, 388–395.
