BIOC 675 Fundamentals of cryo-electron microscopy
Instructors:
Richard Baker (baker@med.unc.edu, GMB 3046)
Joshua Strauss (joshua_strauss@med.unc.edu, Glaxo 008)
Meeting Dates: 01/11/2024 – 02/15/2024
Times: TTh 2:00PM – 3:15PM
Location: GMB 3007
This module will provide a historical and technical overview of cryo-electron microscopy (cryo-EM), with a focus on recent developments in single-particle cryo-EM for high-resolution protein structure determination. The course will provide an overview of current methods in cryo-EM, including a theoretical framework for data collection, data processing, and structure determination. It will consist of eight lectures (1.5 h each), two sessions of hands-on training in the cryo-EM core, homework assignments and an exam.
Fundamental of cryo-EM
- Anatomy of a microscope
- Applications of cryo-EM to biological problems
- Historical perspective of cryo-EM
Image formation, aberrations, and beam-induced motion
- Theory of image formation
- Contrast transfer function (CTF)
- Spherical aberration
- Beam-induced motion and motion correction
Classification, refinement, and reconstruction of 3D models
- 2D and 3D classification
- High-resolution refinement
- Central slice theorem
- Statistical methods in cryo-EM
Sample preparation and practical considerations in cryo-EM
- Cryo grid making
- Air-water interface problem
- Strategies for high-resolution data collection and 3D model refinement
Hands-on workshop for single-particle cryo-EM
- Preparation of cryo grids
- Collection of data
- Determination of a high-resolution structure