COURSES @ CRG: Foerster Resonance Energy Transfer (FRET) microscopy as a tool for functional imaging and quantitative analysis

Organizers: Timo Zimmermann (CRG, Barcelona, Spain); Hernán López-Schier (Helmholtz Zentrum München, Germany)
 
 
Course Overview:

FRET measurements provide the possibility to detect and to quantify biological processes at the molecular level. The use of highly specific biosensors allows to observe events like phosphorylation, Ca2+ signaling and protein cleavage in vitro as well as in fixed or living biological samples. The workshop aims to offer an overview of the state of the art in probe design and the currently available imaging and analysis methods for FRET-based sensors.
  

Teachers:

Raik Grünberg, University of Montréal CA
Olivier Pertz, University of Basel, Basel CH
Tony Squire, Biomedicum, Helsinki FI
Peter Verveer, MPI of Molecular Physiology, Dortmund DE

Scientific Summary of the Course:

The microscopic detection methods (and the subsequent analysis) as well as the development of increasingly sensitive and specific biosensors have made enormous progress in the last years and continue to be areas of highly active development. Especially the use of fluorescent proteins as genetically encodable reporters for molecular interactions and intramolecular conformation changes allows the study of molecular events in real time in living cells, tissues and animals.

Almost any development in fluorescence microscopy allows the improvement of already established FRET detection methods. This can be by faster or more sensitive imaging or by the application of new imaging architectures like Single Plane Illumination Microscopy (SPIM). Lifetime imaging methods that are very well suited for FRET detection are becoming more widely available and offer the potential of fully quantitative FRET measurements that provide full information on the interacting population of molecules.

• probe design
• biological applications examples
• microscope detection methods
• data analysis

The considerations for designing FRET based biosensors will be explained in talks by specialists in this field. The potential for biological experiments will be covered by scientific talks and partially in practical demonstrations and sessions on microscopes on site. An overview of the existing FRET detection methods will be given in seminars and these will be applied in the practical sessions. The analysis of FRET experimental data will be explained theoretically and will be executed in hands-on sessions on analysis workstations under expert supervision.

The limited number of participants in the workshop will allow to provide practical training for each participant in addition to the lectures. For the practical part the whole range of currently commercially available microscope systems will be made available right up to Fluorescence Lifetime Imaging Microscopy (FLIM) by time-correlated single photon counting (TCSPC). Additionally an outlook will be given on using intensity or FRET-based biosensors in organisms using SPIM equipment available in the institute.

A special focus will be put on the analysis of FRET data which will comprise almost one third of the workshop. The participants should therefore come back from this workshop with an understanding of sensor design, the basic ability to perform FRET measurements and the confidence to be able to properly analyze their data, understanding when results are qualitative, when semi-quantitative and when they can be used for global analysis and modeling.

The local organizer and the unit staff have many years of working experience in FRET imaging from basic approaches like acceptor photobleaching and ratiometric Calcium measurements up to the complex analysis required for FLIM experiments. They will be supplemented by a team of expert speakers who are renowned specialists in the fields of probe design, imaging and analysis of FRET data.
  

The number of participants is limited to 25. The positions are open to candidates from institutes worldwide.

Registration deadine was 13/07/2012.
All fields are mandatory. In addition, please upload in the "Abstract" field a document with the answers to the three following questions:

1. What are you currently working on? (up to 1000 characters)
2. Why would this course make a difference in your research? (up to 1000 characters)
3. In case you're selected as a participant of this course, would you need accommodation?