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
