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| 3.5 days 8 – 11 June 2009 Applications are requested from students and post-docs who wish to attend a 3.5 day imaging workshop at UCL focusing on confocal microscopy techniques. The workshop will run from the morning of the 8th June to lunchtime on the 11th June. Registration and accommodation costs will be covered by the ZF-MODELS Integrated Project ([Only registered users see links. ]). Course outline: Day 1 – lectures Basics of microscopy: setting up Koehler illumination, Phase contrast, DIC. Point spread functions and optical resolution. Objectives – what do all the codes mean? What are the benefits and drawbacks of the different objectives? Which one should I choose for what job? Introduction to confocal microscopy: Confocal principle, optical paths, signal cross talk. Digital images, optical vs digital resolution, pixels, voxels and grey levels. Digital and physical regions of interest - scanning and analysis. Optical sectioning and 3D reconstruction and rendering methods. Live cell imaging confocal and multiphoton confocal microscopy: FRET, FRAP, FLIP, FLIM. 2 photon concept, advantages and drawbacks. SHG and THG imaging. Image analysis and 3D reconstruction: Image composition and properties. Signal co-localization analysis. Deconvolution analysis. Application of algorithms to improve image quality – dangers! 3D reconstruction, volume rendering and visualization. Comparison of software packages available. Fluorescent probes and cell labeling: Basics of fluorescence, luminescence and spectral properties and consideration. Chemical fluorophores, fluorescent proteins (standard and advanced), FLAsh ReAsh bi-arsinical probes. Labelling techniques, bathing, AM dyes, electroportation, transfection microinjection, gene gun. Day 2 & 3 & 4 – circus of practical classes to rotate round in groups of 4 (to make sure everyone gets hands on experience). 1. Basic microscope set up and alignment for Koehler illumination, Phase contrast, DIC. Alignment of Hg lamps. Confocal basics – image acquisition, correct dynamics range, eliminate cross talk, Z series, lambda scan, image projection and rotation. 2. Live cell imaging FRAP and FRET techniques. 3. Multiphoton and second harmonic imaging. 4. Image analysis and 3D reconstruction practical. 5. Zebrafish specific module - high-resolution fish brain imaging, including some detail on transgenics, mounting, imaging, reconstruction and presentation Deadline for applications: 10th May 2009 To apply, send a CV & cover letter explaining why you would like to attend and a letter of support from your lab to Steve Wilson ([Only registered users see links. ].uk) |
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