Sample Preparation Workshop

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:

  1. 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
  2. Clip and prepare samples for loading into an Autoloader equipped system
  3. Safely manipulate frozen samples for storage and shipping
  4. Visually identify basic issues arising from sample preparation and propose fixes

Optionally, with another day, 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.

 


 

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