Mitigating risk and enhancing reproducibility in cryo-EM sample preparation

Abstract

Since the invention of the blot and plunge vitrification protocol by Jacques Dubouchet and his colleagues 40 years ago, very little has changed in the basic technique. While the blot and plunge formula has unlocked a world of scientific discovery, the technique is lagging the innovations and scale facilitated by new detectors, software and microscope technology and often comes with a high risk of failure. Early pioneers of the technique are themselves calling for more research to develop alternatives to blotting, positing the fundamental microfluidic physics involved in the traditional model produce inconsistent ice and other negative effects on samples that are ultimately out of the investigator's control. New automated grid vitrification devices are overcoming the limitations of traditional blotting physics while simultaneously making cryo-EM more accessible to researchers of any skill level. We highlight some of the work of our academic partners made possible with a novel alternative to blot and plunge, utilizing pin-printing and jet-vitrification and how researchers have achieved new levels of productivity and predictability using less volume of sample than ever before.