The Kuwait Advanced Microscopy Workshop On Laser Confocal Microscopy March 21-25, 2010 Day 2 22 March 2010 Lecture 4 (LCM): Fundamental techniques behind confocal microscopy: Basics and advance techniques in laser scanning confocal microscopy, image formation, 3-D reconstruction and a choice of microscopic methods will be discussed. Practical 4 (LSM): Optical sectioning in LSM: Sequential and simultaneous collection, advance imaging, screen averaging, kalman averaging, collecting to peak and exponential collection will be practiced. Day 3 23 March 2010 Lecture 5 (LSM) The process of fluorescence; photo bleaching, the selection of appropriate dyes and filters; Immuno-fluorescence labelling (e.g., single, double and triple labelling) will be covered. Practical 5 (LSM): Preparation of frozen and fixed samples; a demonstration of correct controls for double or triple labeling: Background control, bleed- through control for double or triple-labeled samples. Lecture 6 (LSM) Live cell and maintaining healthy cells for live cell imaging (e.g., cell structure and integrity, various imaging chambers, temperature control, pH control, Buffers, O2, CO2 levels will be covered. Fluorescent Probes (e.g., Organelle probes, Membrane probes, probes for nuclear stains and tracers) will be discussed. Practical 6 (LSM): Live cell labelling and collection of XYT, XYZT: Using thin epithelium on the inner surface of the onion layer and staining with various organelles probes live motion will be collected in time laps.
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