Morning session

10.00-11.00         Basics of microscopy (Martin Spitaler, FILM)
magnification, properties of light, resolution, transmitted light contrast (PH, DIC, darkfield), epifluorescence, confocal microscopy

11.00-11.45         Live imaging techniques (Mark Scott, FILM)
Multiphoton, in vivo imaging, second harmonic generation (SHG), photobleaching, phototoxicity

11.45-12.00         coffee & tea break

MICROSCOPY MATCH-MAKER

12.000-13.00        Microscopy Match-Maker:
FILM users present their research performed in FILM through 3 min snapshot presentations

13.00-14.00         Sandwich lunch

Afternoon session

14.00-14.45         A walk through the zoo: Molecular  imaging tools (Martin Spitaler, FILM)
FRAP, FLIP, FLIM, FRET, FCS, FUEL, BRET and more

14.45-15.30         Super-resolution microscopy (Steve Rothery, FILM)
TIRF, PALM, STORM

15.30-15.45         coffee & tea break

15.45-16.30         Understanding images (Luis Pizarro, Computing)
basics of image data analysis, like noise removal, segmentation, tracking, registration

PLENARY LECTURE

16.30-17.30         Plenary talk:
                                Spencer Shorte (Institute Pasteur, Paris):
                                "FUEL for thought: a new method for detection of spatial proximity
                                  on mesoscopic scales in vitro and in vivo using luminescence excitation"

Bioluminescence imaging is a powerful technique facilitating a broad range of applications from cell-based assay to in vivo analysis in living, intact organisms. The current state of the art uses Bioluminescence Resonance Energy Transfer (BRET) to improve the sensitivity of bioluminescence by red-shifting blue photons, and providing a measure of molecular co-localization at distances of up to 10nm. However, BRET detection methods may overlook long-distance, radiating energy excitation-emission effects that are significant in the bioluminescent detection regime (1). Fluorescence by Unbound Excitation from Luminescence (FUEL) describes this radiating luminescence effect that excites fluorophores by epifluorescence at distances far beyond 10nm, many microns, or even millimeters away in a manner completely distinct from BRET. Further, we show that detection of FUEL per se is sufficient to provide a detection of long-distance proximity in, and beyond the microscopic range both in vitro and in vivo. By enabling detection of mesoscopic proximity between luminescent and fluorescent probes in the context of living biological tissues FUEL promises utility as a novel tool for high-content analysis in cell and animal models.

1. Dragavon J, Blazquez S, Rekiki A, Samson C, Theodorou I, Rogers KL, Tournebize R, Shorte SL. In vivo excitation of nanoparticles using luminescent bacteria. Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):8890-5.

The plenary lecture will be followed by refreshments