Cryo-EM Sample Preparation

This protocol describes the steps for preparing cryo-EM samples.

1. Grow cells to mid-log phase.
   • Culture cells in appropriate media to a density of 1-2 x 10^6 cells/mL.
2. Harvest and wash with buffer.
   • Centrifuge cells and resuspend in a suitable buffer (e.g., PBS or Tris-HCl).
3. Apply sample to grids and plunge freeze.
   • Apply 3-4 µL of the sample to a glow-discharged cryo-EM grid.
   • Blot away excess liquid and rapidly plunge-freeze in liquid ethane.