Sample Preparation (Sept. 2022)

Length: 3 days

Dates: Sept. 20-22nd

Cost: No cost

Lodging: Room and Board provided

Trainees: 4-6 Trainees

Who: PNCC Awarded General Access Proposal members only

Location: RLSB, 2730 S.W. Moody Ave, Portland, OR 97201

Accommodations: University Place Hotel

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 September 20th through 22nd, 2022 with the 22nd 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.

NOTE: Applicant must be on an active General Access Proposal. PNCC prefers to train one lab member per group with the intention that this member can further train other members of their group.

 

Agenda:

From 9am to 5pm (Pacific) each day:

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