Frozen-hydrated rotavirus double-layered particles

Methods

frozen-hydrated rotavirus double-layered particles (0.01 mg/mL concentration, 2 µL volume)
prepared on Affinity Grids decorated with His-tagged protein A and antibodies against the outer capsid protein, VP6
specimen prepared using Cryoplunge™ 3 instrument with GentleBlot™ technology on C-flat™ 2/1 grids, Protochips, Inc.
specimens examined under low dose conditions at 120 kV using a 626 liquid nitrogen cryo transfer holder
recorded at 50,000x, dose of  ~5 electrons/Ų