Leica EM GP2 Standard Operating Procedure
1. Purpose
Setting up, operating, and shutting down the Leica EM-GP2 in a safe and effective manner for the preparation of frozen cryo-EM of samples. Note: This SOP is for BSL-1 work only.
2. Definitions:
2.1 Leica GP2 is a vitrification device for freezing specimens to use for cryoEM analysis.
2.2 Liquid Nitrogen (LN2) is a cryogenic liquid stored under pressure.
2.3 Ethane is classified as a flammable gas.
3. Supplies & Equipment
- PPE (BSL-1)*(also see site specific instructions)
- Laboratory Coat
- Nitrile Gloves
- Goggles / Safety Glasses
- Cryogenic Gloves
- Face Shield
- Chemicals/Reagents
- Liquid Nitrogen
- Ethane
- 70% Ethanol
- Distilled Water
- Sample
- Filter Paper (Pre-punched Whatman #1 or #541)
- Pipette and Tips
- Tweezers for misc. grid handling
- Leica GP2 Tweezers
- Glow Discharged / Plasma Cleaned EM Grids
- Grid Storage Boxes/Buttons
- Grid Box Lid Opening Tool (Screwdriver or TFS Pin Tool)
- LN2 Transfer Dewars
- Forceps (fine tip and long tweezers)
4. Procedure
4.1 Instrument setup and cryogen preparation (takes 30-60 min) *(also see site specific instructions for 4.1)
4.2 Freezing Cycle / Plunge Freezing
4.3 Shutdown *(also see site specific instructions for 4.3)