Dear microscopists, here the latest update on today's FILM Microscopy Day, including *Spencer Shorte's invited talk: "FUEL for thought: a new method for detection of spatial proximity on mesoscopic scales in vitro and in vivo using luminescence excitation"* (4.30pm, G34, see abstract below) and the list of short presentations during the Microscopy Match Maker. See you there, Martin ------------------------------------------------------------------------ *FILM MICROSCOPY DAY 2013** **Friday 28 June 2013** **G34** *** ------------------------------------------------------------------------ PROGRAMME: ------------------------------------------------------------------------ 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 *Alejandra Garcia-Cattaneo* Ajuba is a key component for adherens junction maintenance *Ciro Chiappini* Combining Confocal and Electron Microscopy to investigate the cell-nanoneedle interface *Edwin Hawkins* *Jill Johnson* 3D imaging of airway remodelling in asthmatic mice *Kevin Woollard* Study of in-vivo monocyte and macrophage biology by intravital microscopy *Kim Jonas* The Use of simultaneous dual colour PALM for imaging of receptor complexes *Reema Khorshed* Computational analysis of in vivo images *Ryan Robinson* Ca2+ Imaging of Guinea Pig Ganglia Neurons -- Investigating the effects of Diesel Exhaust Particles (DEP) *Sean Warren* FLIMFit: Rapid Global Analysis of FLIM Microscopy Data *Sophie Schobesberger* *Sowmya Purushothaman* *William Jackson* Tracking blood monocyte motility in vitro *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 ------------------------------------------------------------------------ *Martin Spitaler, PhD* *FILM - Facility for Imaging by Light Microscopy* - Facility Manager - Sir Alexander Fleming Building, desk 401 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK Tel. +44-(0)20-759-42023 E-mail m.spitaler@imperial.ac.uk <mailto:m.spitaler@imperial.ac.uk> Website: http://imperial.ac.uk/imagingfacility