Module 4: DNA organization in eukaryotes and bacteria
Organisers
M. Nollmann, Centre de Biochimie Structurale, CNRS. See website.
Aims
The aim of this module is to demonstrate the use of super-resolution methods to investigate nuclear architecture in eukaryotes and bacteria, and what factors are determinant in establishing/ regulating nuclear architecture. Three super-resolution methods will be approached: (1) Structured illumination Microscopy; (2) single-molecule tracking; and (3) STORM/ PALM. Experts in each of these fields will give talks in which they will provide concrete examples on how to use these problems to solve concrete biological questions. Practical sessions will provide hands-on experience to using 3D-SIM, and single-molecule tracking/PALM to investigate the structures and dynamics of proteins involved in defining chromosome structure. An image analysis session will be devoted to teach students to properly analyse images, and to pinpoint typical reconstruction/image analysis problems.
Lectures
Lectures of the module will by given by:
Practical Sessions
M. Nollmann, Centre de Biochimie Structurale, CNRS. See website.
Aims
The aim of this module is to demonstrate the use of super-resolution methods to investigate nuclear architecture in eukaryotes and bacteria, and what factors are determinant in establishing/ regulating nuclear architecture. Three super-resolution methods will be approached: (1) Structured illumination Microscopy; (2) single-molecule tracking; and (3) STORM/ PALM. Experts in each of these fields will give talks in which they will provide concrete examples on how to use these problems to solve concrete biological questions. Practical sessions will provide hands-on experience to using 3D-SIM, and single-molecule tracking/PALM to investigate the structures and dynamics of proteins involved in defining chromosome structure. An image analysis session will be devoted to teach students to properly analyse images, and to pinpoint typical reconstruction/image analysis problems.
Lectures
Lectures of the module will by given by:
- Specific-module lecture 1: Maxime Dahan (Institut Curie, Paris): Target search of DNA-binding proteins in mammalian cells. Tue 11am.
- Specific-module lecture 2: Melike Lakadamyali (ICFO, Barcelona): STORM imaging to investigate chromosome architecture in human cells. Wed 10:45.
- Specific-module lecture 3: Everything you ever wanted to know about 3D-SIM and never dared asking. Short talks by Lothar Schermelleh and Antoine Le Gall (CBS, CNRS, France). Wed 20/01 at 11am.
- Specific-module lecture 1: Nitty Gritty of single-molecule localization microscopy. Short talks and discussion with Mat Stracy, Jeff Moffitt, and JB Fiche (CBS, CNRS, France). Thu 21/01 at 11am.
Practical Sessions
- Session 1: 3D-SIM imaging
- Session 2: 3D STORM/PALM imaging using adaptive optics
- Session 3: spt-PALM