Cryoelectron Microscopy of Macromolecular Assemblies

Cryo electron microscopy in combination with image analysis is increasingly powerful in producing 3D structures of individual molecules and large macromolecular complexes that are unapproachable by other methods.

This comprehensive course will cover the theory and practice of solving molecular structures by electron microscopy. We will start with optics, sample preparation and a basic mathematical description of diffraction before moving into a detailed exploration of the three main methods of structure determination: tomography, single particle analysis, and 2D crystallography. We will end with a discussion of map interpretation and molecular fitting.

This course will have two components

• A series of lectures from local experts followed by student-led discussion sessions.
• A series of practical sessions that parallel the topics introduced during the lectures.

Both components will be held on the premises of the New York Structural Biology Center.

Students and postdocs from all NYC institutions are encouraged to attend
Attendance to the practical sessions will be limited and preference will be given to students enrolled for credit

Textbooks relevant for the course are

• "Three Dimensional Electron Microscopy of Macromolecular Assemblies" by Joachim Frank
• "Electron Crystallography of Biological Macromolecules" by Glaeser, Downing, DeRosier, Chiu

Course Director: David Stokes 212-263-1580 stokes@saturn.med.nyu.edu

The New York Structural Biology Center is located on the south campus of City College
89 Convent Ave at 133rd St in Manhattan
A,C,B,D trains to 125th St or 1 train to 137th St

Registration Information

Please notify the course director (David Stokes) of your intention of taking the course.

For graduate students wishing to take the course for credit:

This course will be listed in the catalogue of the New York University School of Medicine (http://www.med.nyu.edu/sackler/courses).

listed as G16.4408 - Cryoelectron Microscopy of of Macromolecular Assemblies.
Call # 31569
3 credits for the lecture/discussion
1 credit for the practical sessions

• NYU students should enroll through the Sackler Inst.
  
• Mt. Sinai students should enroll through NYU GSAS using the above course information.
  
• Albert Einstein College of Med students should contact Hernando Sosa or Todd Evans.
  
• Rockefeller University students should contact Seth Darst or Sid Strickland.
  
• Students from Wadsworth or SUNY Albany should contact Carmen Mannella.
  
• Students from Columbia or CUNY can enroll through the Inter-University Doctoral Consortium
  
• Other students should contact the Dean of Graduate Studies or your institutional representative on the NYSBC CryoEM Operations Committee to learn about procedures for taking this course for credit.
  

• practical on negative stain sample prep - Sept 20

Optics, waves, image formation - Sept 25 - Ruben Diaz (NYSBC)

• practical on frozen-hydrated sample prep - Sept 27

Diffraction and Fourier transforms - Oct 2 - Iban Ubarretxena (CUNY)

• practical on optical and computed FFT - Oct 4

Fundamentals of Image Analysis - Oct 9 - Bill Rice (NYSBC) practical on digital imaging - Oct 11

Tomography I - Oct 16 - Carmen Mannella (Wadsworth Ctr)

• practical on tomography I (IMOD reconstruction) - Oct 18

Single Particle Analysis I - Oct 23 - Joachim Frank (Wadsworth Center)

• practical on tomography II (IMOD segmentation) - Oct 25

Single Particle Analysis II - Oct 30 - Joachim Frank (Wadsworth Center)

• practical on single particle analysis I (particle picking and classification) - Nov 1

Tomography II - Nov 6 - Carmen Mannella (Wadsworth Ctr)

• practical on single particle analysis II (refinement & reconstruction) - Nov 8

Electron Crystallography - Nov 13 - Iban Ubarretxena (CUNY)

• practical on single particle analysis III (multivariate statistical analysis) - Nov 15

Helical reconstruction - Nov 20 - Hernando Sosa (Albert Einstein College of Medicine)

• practical on 2D crystallography (2DX) - Nov 22

Hybrid Methods - Nov 27 - David Stokes (NYU/NYSBC)

• Thanksgiving - Nov 29

Molecular fitting - Dec 4 - Hernando Sosa (Albert Einstein)

• practical on molecular visualization (Chimera) - Dec 6