Fig. 3 |. Infrastructure for anaerobic single-particle cryoEM grid preparation.

a, Ethane/propane is dispensed within a 50 mL Falcon tube on liquid nitrogen. b, The Vitrobot Dewar is assembled and placed within a secondary Dewar to capture any spilled cryogen. Before bringing into the tent, the outer reservoir is filled with liquid nitrogen. c, All Dewars to be brought into the tent containing cryogen are covered with aluminum foil secured with tape. d, Customized vinyl anaerobic tent containing a ThermoFisher Vitrobot Mark 4. e, Copper tubing is threaded out of the anaerobic tent to a vacuum pump, which vents to the fume hood during grid preparation. f, Schematic overview of setup for anaerobic single-particle cryoEM grid preparation including illustration of customized ports for an Amicon stir cell for protein concentration and the port for atmospheric exhaust during grid preparation.