October 7-10, 2024
Instruct Hands-on Workshop
Sample Preparation for Single Particle Analysis by Cryo-Electron Microscopy
Braga - Portugal
Braga - Portugal
Funded by:
Organized by:
Sponsored by:
About the Workshop
The workshop aims to teach participants the best practices in sample preparation methods for cryo-EM imaging and introduce them to the image data acquisition workflow. The course will include lectures on single particle analysis (SPA), with special focus given to tutorials/practical sessions on the preparation of biological samples for negative staining and quality assessment, vitrification on a plunge-freezing device, and grid optimization/screen of samples with a cryo-transmission electron microscope (TEM). Participants are encouraged to bring their own samples.
The most recent advances in protein production and stabilization and the most recent technological advancements for grid preparation and sample manipulation will also be highlighted and discussed, providing a complementarity view to achieve truly integrative structural biology approaches to tackle challenging scientific projects.
At the end of the course, students will have a firm grasp of the individual aspects of sample preparation for SPA by cryo-EM. They will be able to develop a holistic vision of the integrated use of biophysics, biochemistry, and cryo-EM for protein structure determination and analysis.
Join this unique opportunity!
Organizing Committee:
José A. Brito (ITQB NOVA)
Margarida Archer (ITQB NOVA)
Oliver Schraidt (INL)
Paulo Ferreira (INL)
Jose Maria Carazo (CNB–CSIC)
Who should attend?
The workshop aims to teach participants the best practices in sample preparation methods for cryo-EM imaging and introduce them to the image data acquisition workflow is ideal for researchers new to cryoEM, especially PhD students and early postdocs.
Focus: Dive deep into single particle analysis using cryoEM.
Curriculum Highlights:
Basic theory and applications
Protein production
Sample preparation
EM technologies
Intimate Learning Environment: Limited to 12 participants for personalized interaction with tutors and speakers.
Application Requirements:
Short CV
Motivation letter
Support letter from Supervisor
Deadline for application: August 19, 2024
Jumpstart your cryoEM journey with hands-on training and expert guidance!
Venue
This workshop will be held at the INL - International Iberian Nanotechnology Laboratory, Braga, Portugal. INL was established in 2005 by a joint decision of the Governments of Portugal and Spain and is an international intergovernmental research organization aiming to perform cutting-edge research and development in nanotechnology and to function as an innovation integrator in multiple application domains.
Operating since late 2010, INL infrastructure provides a high-tech research environment for INL staff and users. The Advanced Electron Microscopy, Imaging & Spectroscopy (AEMIS) facility is a core multi-user facility with cutting-edge instrumentation, techniques, and expertise required to characterize physical and life sciences samples.
INL hosts a Glacios 200kV machine with Falcon 4i detector that is used for SPA, tomography and MicroED. The Glacios features a MicroED package with Ceta-D camera and has the Multigrid feature, smart EPU and Athena installed. Negative stain samples are checked with a JEOL 2100 microscope and sample vitrification is performed with either a ThermoFisher Vitrobot Mark IV or a Leica Plunge Freezer EM GP2.
Programme
DAY 1 | October 7, 2024
09h00 Welcome, Paulo Ferreira (INL)
09h15 Course Outline and General Information, José A. Brito (ITQB NOVA)
09h30 Fundamentals of Electron Microscopy, Jose Maria Carazo (CNB–CSIC)
10h30 Coffee break
11h00 Cryo-EM Techniques and Applications - An Overview, Oliver Schraidt (INL)
12h00 Sponsored Talk, Thermo Fisher Scientific
13h00 Lunch
14h30 Practical Sessions (in parallel)
Session 1 Negative Staining (Group 1), José A. Brito (ITQB NOVA) & Ana Malheiro (INL)
Session 2 Plunge Freezing, Clipping and Loading (Group 2), Marta Carroni & Dmitry Semchonok
Session 3 Grid Screening at Glacios Cryo-TEM and Preliminary Data Analysis (Group 3), Oliver Schraidt (INL) & Jose Maria Carazo (CNB–CSIC)
17h30 Ice break Session & INL Tour
20h00 Social Dinner
DAY 2 | October 8, 2024
09h00 The Portfolio of Instruct-ERIC for Structural Biology Studies, Margarida Archer
09h30 Principles of Cryo-EM Data Processing, 2D & 3D Classification, Jose Maria Carazo (CNB–CSIC)
10h30 Coffee break
11h00 Sample Preparation for Cryo-electron Microscopy: From Negative Staining to Single Particle Analysis, Marta Carroni
12h00 Science-Case 1, João Morais Cabral
13h00 Lunch
14h30 Practical Sessions (in parallel for Groups 1, 2 and 3)
17h30 Poster Session 1 (odd numbers)
DAY 3 | October 9, 2024
09h00 Sample Quality Assessment and Improvement for Downstream Structural Studies: A Biophysical and Biochemical Approach, José A. Brito (ITQB NOVA)
09h45 3D cryo-EM map Calculation, Resolution Estimation, Model Building and Refinement, Dmitry Semchonok
10h30 Coffee break
11h00 “Structural basis for bacterial protein disaggregation and proteolysis”, Marta Carroni
12h00 Sponsored Talk
13h00 Lunch
14h00 Practical Sessions (in parallel for Groups 1, 2 and 3)
17h00 Poster Session 2 (even numbers)
DAY 4 | October 10, 2024
09h00 Cryo-EM in Industry, BluePharma
09h30 CryoSPARC – capabilities and new features, Stavros Azinas
10h30 Coffee break
11h00 CryoSPARC – capabilities and new features (cont), Stavros Azinas
12h00 Q&A session
13h00 Lunch
14h00 Departure
Registration
ACADEMIC
Registration €250,00 for an all-inclusive package.
What’s Covered?
Hands-on practical sessions
Lunches
Coffee-breaks
Social Dinner
Comfortable accommodation for 4 nights (shared double-rooms)
Networking Opportunities: Share experiences and insights with peers and experts during breaks and poster sessions.
Comprehensive training.
Secure your spot and make the most of this incredible learning opportunity!
NON-ACADEMIC
Registration €500,00 for an all-inclusive package.
What’s Covered?
Hands-on practical sessions
Lunches
Coffee-breaks
Social Dinner
Comfortable accommodation for 4 nights
Networking Opportunities: Share experiences and insights with peers and experts during breaks and poster sessions.
Comprehensive training.
Secure your spot and make the most of this incredible learning opportunity!
Let’s talk!
For any queries, don't hesitate to get in touch with ourTeam.
Please fill in the form, and we will be in touch soon!
INL – International Iberian Nanotechnology Laboratory
Av. Mestre José Veiga
4715-330 Braga | Portugal