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Instructors:

Richard Baker (baker@med.unc.edu, GMB 3046) 

Joshua Strauss (joshua_strauss@med.unc.edu, Glaxo 008) 

Meeting Dates: 03/28/2022 – 4/27/2022

Times: TTh 2:00 – 3:00pm

Location: GMB 3007

BIOC 675 cryoEM Syllabus

 cryoEM Core Facility 

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