Abstract Jet vitrification harnesses the exceptionally high cooling potential of rapidly moving liquid cryogens to achieve ultrafast sample freezing with superior cooling efficiency. Building on earlier work, this study provides a comprehensive investigation into the physical and technical principles underlying dual-jet vitrification. Developing ultra-rapid freezing methods that reliably produce vitreous cells, supports thicker samples, and integrate seamlessly into cryo-electron tomography (cryo-ET) workflows do represent important steps forward in the field. Here, we introduce an experimental platform designed to study the dynamics and efficacy of liquid-ethane jet vitrification at −180 °C. The system integrates multiple modules enabling controlled examination of the vitrification process at microsecond timescales, with a focus on quantifying temperature transitions, identifying optimal freezing parameters, and delineating the limitations of dual-jet configurations. The compact design allows operation within a standard biosafety cabinet and supports various sample holders compatible with biological specimens. Furthermore, we demonstrate how this setup can be directly incorporated into existing cryo-ET workflows using EM grids pre-mounted in AutoGrids. Overall, this work establishes a versatile platform for dissecting the mechanisms of ethane jet vitrification and provides methodological insights that support the preparation of thicker, well-preserved biological samples for in situ structural analysis.
Premaraj et al. (Wed,) studied this question.