Workshop for Cryo-electron Tomography 10-14 March 2025

This four and half day workshop will cover cryo-ET workflows from sample preparation on EM grids to data processing and analysis.

Application

Speakers

Program

Contact

OVERVIEW

Workshop description

Cryo-electron tomography (cryo-ET) is a powerful structural technique. Its power comes from its versatility to study biological systems in vitro or in situ, i.e. within cells or tissue. It has the ability to reach uncharted regions in biological systems at high resolution. Advancements in microscopes, cameras, and computation make it now possible to determine sub-10 Å 3-dimensional structures of molecules directly within cells.

This four-and-a-half-day workshop will cover cryo-ET workflows from sample preparation on EM grids to data processing and analysis. The workshop will be a mixture of lectures and practical sessions. Participants will get hands-on experience in sample preparation on EM grids (micropatterning), sample freezing (plunge freezing and high-pressure freezing), cryo-correlative light and electron microscopy (cryo-CLEM), cryo-focused ion beam milling (cryo-FIB), cryo-TEM tiltseries data collection, and data processing and analysis (MotionCor2, IMOD, Dynamo).

It is compulsory that applicants have previous cryo-EM experience either with single-particle or already with cryo-tomography.

This workshop is supported by SciLifeLab, CryoNET and Umeå University and organized by the Umeå Centre for Electron Microscopy (UCEM).

Workshop organisers

Head organizer: Erin Schexnaydre
Assistant organizers: Tanvir Shaikh, Sara Sandin and Lorene Gonnin

Entry Requirements

Priority will be given to researchers involved in a project where the use of cryo-ET is needed, which are to be briefly described upon application. The workshop is suitable for researchers in the life sciences with a master's or PhD degree, including graduate students, postdocs and staff scientists.

APPLICATION

Apply here latest by Monday, January 13th, 2025

* this information is required

First Name *

Last Name *

E-mail *

Position *

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Dietary requirements

Level of experience with cryo-EM/ET *

What makes your project applicable for cryo-ET? *

Please describe your motivation for participating in this workshop *

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SPEAKERS

Speakers

Veijo Salo
Postdoctoral Fellow
EMBL Heidelberg
Julia Mahamid’s group

Elisa Lisicki
PhD Student
Max Planck Institute of Molecular Physiology, Dortmund
Stefan Raunser’s group

Oda Helene Schiøtz
PhD Student
Max Planck Institute of Biochemistry, Munich
Jürgen Plitzko’s group

Dustin Morado
Research Engineer
SciLifeLab National Cryo-EM Facility
Stockholm University

Erin Schexnaydre
UCEM Facility Manager
CryoET/cryoFIB/cryoCLEM Specialist
Umeå University

Sara Sandin
UCEM Facility Manager
CryoEM Head of Unit
Umeå University

Tanvir “Tapu” Shaikh
UCEM Facility Manager
Image Processing Specialist
Umeå University

Marie Sorin
Postdoctoral Fellow
Department of Medical Biochemistry and Biophysics
Umeå University

Lorene Gonnin
Postdoctoral Fellow
Department of Chemistry
Umeå University

PRELIMINARY PROGRAM

Downloadable program (PDF)

Day 1 Morning (8:45-13:00)

08:45-09:00 Coffee (Glasburen)
09:00-09:15 Welcome and course expectations (Glasburen, Speaker: Erin Schexnaydre)
09:15-10:15 Coffee break and project discussion (Glasburen)
10.15-11:00 Cryo-ET Research Talk (Glasburen, Speaker: Marie Sorin)
11:00-12:00 Cryo-ET sample preparation Part I (Glasburen, Speaker: Lorene Gonnin)
12:00-13:00 Lunch

Day 1 Afternoon: Practical Sessions (13:00-17:00)

Group 1 Vitroblot plunge freezing (UCEM facility)
Group 2 High pressure freezing (C2)
Group 3 Micropatterning (A6 44 15)
14:30-15:00 Coffee break at UCEM facility

Day 2 Morning (9:00-13:00)

09:00-10:00 Sample preparation part II (Glasburen, Speaker: Erin Schexnaydre)
10:00-10:15 Coffee Break (Glasburen)
10:15-11:00 Cryo-CLEM research talk (Glasburen, Speaker: Veijo Salo)
11:00-11:45 Cryo-Lift out (Glasburen, Speaker: Oda Schiøtz)
12:00-13:00 Lunch

Day 2 Afternoon: Practical Sessions (13:00-17:00)

Group 1 Cryo-CLEM (UCEM facility)
Group 2 Cryo-FIB-SEM (UCEM facility)
Group 3 Cryo-TEM (UCEM facility)
14:30-15:00 Coffee break at UCEM facility

Day 3 Morning (9:00-13:00)

09:00-10:00 Tiltseries data collection (Glasburen, Speaker: Sara Sandin)
10:00-10:15 Coffee break (Glasburen)
10:15-11:00 Tomogram Visualization and analysis (Glasburen, Speaker: Tapu Shaikh)
11:00-11:45 Research talk (Glasburen, Speaker: TBA)
12:00-13:00 Lunch

Day 3 Afternoon: Practical Sessions (13:00-17:00)

Group 1 Cryo-TEM (lamella) (UCEM facility)
Group 2 Cryo-FIB-SEM (lift-out) (UCEM facility)
Group 3 Data reconstruction and analysis (Computer lab)
14:30-15:00 Coffee break at UCEM facility

Day 4 Morning (9:00-13:00)

09:00-10:00 Subtomogram averaging (KBF301, Speaker: Dustin Morado)
10:00-10:15 Coffee break (KBF301)
10:15-12:00 STA Practical (Computer lab)
12:00-13:00 Lunch

Day 4 Afternoon: Practical Sessions (13:00-17:00)

Group 1 Data Reconstruction (Computer lab)
Group 2 Data Analysis (Computer lab)

Day 5 (9:00-12:00)

09:00-10:00 Research Talk (Glasburen, Speaker: Elisa Lisicki)
10:00-10:15 Coffee break
10:15-12:00 Practical continuation and Project discussions (Computer lab)