Length: 3 days
Dates: Postponed due to Covid-19, New Dates TBD
Cost: no cost
Lodging: Room and Board provided
Trainees: 4-6 Trainees
Who: PNCC Awarded Proposal members only
Location: RLSB, 2730 S.W. Moody Ave, Portland, OR 97201
Pacific Northwest Center for Cryo-EM (PNCC) would like to offer a three-day hands-on training workshop for 4-6 trainees on “Introduction to basic theory and hands-on practice in preparing cryo-EM specimens for SPA”. This short course will be held at PNCC between January 21-23rd, 2020. Students do not require any pre-existing knowledge of cryo-EM specimen preparation. Previous experience with negative-stain sample preparation may be useful but is not required. At the end of this module students will:
- Be able to successfully freeze one of three benchmark samples using one (or more) of the following PNCC devices: TFS Vitrobot Mk IV, Leica GP2
- Clip and prepare samples for loading into an Autoloader equipped system
- Safely manipulate frozen samples for storage and shipping
- Visually identify basic issues arising from sample preparation and propose fixes
Optionally, with another day, students may also:
- Practice using their own samples with feedback from PNCC trainers
- Practice using different grid substrates, such as Lacey Film, AuFoil, thin-carbon, and graphene-oxide.
Hands-On Session:
• Basic Cryo Grid Handling and Safety
• Cryo Grid Storage and Shipping
• Autogrid Clipping
Choose 1-2 of:
• TFS Vitrobot Mk IV
• Leica GP2
Hands-On Session: 2-3 Days, Group size 3 (max) per tool, Each user should get 4-6 hours of time practice time
Day 1
08-09: Onsite Badging and Introductions
09-11: Demonstration by PNCC staff: freezing, clipping, handling, storage
11-13: Supervised user practice, at room temperature
13-14: Lunch Break
14-16: Supervised User practice under cryogenic conditions
16-18: Group screening and grid evaluation (one microscope per group)
Day 2
09-11: Demonstration by PNCC staff
11-13: Supervised user practice, room temperature
13-14: Lunch Break
14-16: Supervised user practice under cryogenic conditions
16-18: Final Evaluation, user’s choice of 1 tool/sample.
Optional Day 3
09-13: Users freeze their own samples with SPOC assistance and screening
13-14: Lunch Break
14-18: Users freeze their own samples with SPOC assistance and screening