Sample Preparation

A on-site hands-on training course covering sample vitrification on Vitrobot and Leica GP2.

Dates:  Mar. 7-9th

Lodging: Room and Board provided

Accommodations: University Place Hotel

Location: RLSB

Who: PNCC Awarded General Access Proposal members only

The Pacific Northwest Center for Cryo-EM (PNCC) is offering 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 Single Particle Analysis”. This workshop short course will be held at PNCC from March 7th through 9th, 2023 with the 9th being an optional day. 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. By workshop conclusion, trainees will be able to:

1. Successfully freeze one (or more) benchmark samples using one (or more) of the following PNCC devices: TFS Vitrobot Mk IV, Leica GP2
2. Clip and prepare grids for loading into an Autoloader equipped system
3. Safely manipulate frozen grids for storage and shipping
4. Visually identify vitrification issues and propose fixes

Optionally, on day three, students may also:

1. Practice using their own samples with feedback from PNCC trainers
2. Practice using different grid substrates, such as Lacey Film, AuFoil, thin-carbon, and graphene-oxide.

Agenda

From 9am to 5pm (Pacific) each day.
Hands-On Sessions, Group size 3 (max) per instrument.
Each user should get 4-6 hours of time practicing:

Day 1

• Onsite Badging and Introductions
• Demonstration by PNCC staff: freezing, clipping, handling, storage
• Supervised user practice, at room temperature
• Supervised User practice under cryogenic conditions

Day 2

• Groups Alternating Instruments:
  • Supervised user practice under cryogenic conditions
  • Grid evaluation on screening microscope
• Final Evaluation: Trainees demonstrate mastery of "Sample Preparation Workflow" on either TFS Vitrobot or Leica GP2

Day 3 (Optional)

• Users freeze their own samples with SPOC assistance
• Grid Evaluation on screening microscope