Venue
Remote course
December 14-18, 2020 from 09.30-18.00 CET
Overall aims and course outline
Cryo-Electron Microscopy has established as a mature technique to determine the three-dimensional structure of biological macromolecules. It is currently reaching atomic resolution and it is capable of accessing to the structure of conformationally heterogeneous samples, as well as macromolecular complexes. It is complementary to other structural techniques like X-ray crystallography and Nuclear Magnetic Resonance and at the moment it is the most rapidly growing technique in atomic structure depositions at public databases like PDB.
The aim of the course is to give an overall overview of the whole process of single particle analysis (SPA) starting from sample preparation, image acquisition at the microscope, image processing and atomic modelling. The I2PC has a long tradition of organizing courses of image processing and atomic modelling, with Instruct courses that have been heavily oversubscribed and had to be repeated twice in the same year to be able to attend its demand. In this edition of the course we have included the sample preparation and image acquisition part, which will be done in the recently established high-end CryoEM facility at the CNB, which incorporates a Thermofisher Talos Arctica (200kV) and JEOL CryoARM 300 (300 kV). The CNB CryoEM facility is in the process of being incorporated into the offer of the Instruct catalogue. For this reason, we have extended our traditional 3 days course into a 5 days course that includes sample preparation and image acquisition.
For logistic reasons and travelling restrictions, the whole course will be given online. An additional benefit of this circumstance is that the course can be recorded and stay as a permanent course in Instruct. Although the course is online we will try to keep the interactions with attendees. For this reason, we will limit the maximum number of attendees to 20.
The course is aimed at researchers of all levels wanting to adopt electron microscopy as one of the tools at their disposal for elucidating biological structures.
Expected impact for young researchers
CryoEM is the most rapidly growing structural technique. PhD students as well as young post-docs will certainly benefit from approaching this technique. Knowing it will open new doors to new positions worldwide.
Contacts
Lead contact:
Carlos Oscar Sorzano
email. coss@cnb.csic.es
Administrative Contact:
Blanca Benitez
email: blanca@cnb.csic.es
Tutors
Marta Martínez (MM), Carlos Oscar Sorzano Sánchez (COSS), Roberto Marabini (RM), Roberto Melero (RME), Jose-María Carazo (JMC), José-María Valpuesta (JMV), Javier Chichón (JC)
Program
| Day 1: Data acquisition 9:30 Introduction to the course (JMC, JMV, COSS) 9:35 Introduction to CryoEM (JMV) 10:20 Break 10:30 Introduction to camera detectors (JC) 11:20 Break 11:30 Sample preparation NEGATIVE STAIN (JMV) 12:30 Lunch Break 13:30 Sample preparation VITRIFICATION (JC) 14:20 End of day | Day 2: Data acquisition 9:30 Data Acquisition Lecture 10:20 Break 10:30 Grid Clipping and Microscope loading (JC) 11:20 Break 11:30 Screening 12:30 Lunch Break 13:30 General discussion 14:20 End of day |
| Day 3: Data processing 9:30 Movie alignment, Screening micrographs and CTF (COSS) 10:20 Break 10:30 Particle picking and screening (COSS) 11:20 Break 11:30 2D classification (COSS) 12:30 Lunch Break 13:30 Personal practice 16:30 General discussion 17:00 End of day | Day 4: Data processing 09:30 Initial volume (COSS) 10:20 Break 10:30 Alignment, classification, and reconstruction (COSS) 11:20 Break 11:30 Validation and analysis (COSS) 12:20 Lunch Break 13:30 Personal practice 16:30 General discussion 17:00 End of day |
| Day 5: Data modelling 09:30 Homology modelling (MM, RM) 10:20 Break 10:30 Model alignment and refinement (MM, RM) 11:20 Break 11:30 Model validation (MM, RM) 12:20 Lunch Break 13:30 Model validation (MM, RM) 14:30 General discussion; Dissemination: Infrastructure access through Instruct and iNext-Discovery (RME, JMC, COSS) 14:30 End of day work* | * Virtual machines and I2PC staff will be available until 17:30 for those who want to access. |
Key dates:
October 26 – November 20, 2020: Submission of the course application
23– 30 November, 2020: Communication of awarded applications
REGISTRATION
Registration will be free for all attendees.
Fill your application here. (Registration is now closed)
This course is funded by Instruct – ERIC, an Integrated Structural Biology Infrastructure for Europe and CRIOMECORR Project
