Why use fluorescence microscopy?
Cellular events are accomplished by the coordinated interaction of cellular components within the three dimensional context of a cell. Simultaneous observation of multiple components in three dimensions is therefore essential for understanding such interactions on a cell-by-cell basis. Fluorescence microscopy allows the use of multiple fluorescent markers used to stain specific structures or molecules within a single cell, their three-dimensional acquisition and visualization to analyze the spatial distribution of multiple proteins and nucleic acids in relation to well-defined cellular or nuclear structures, a resolution of the order of 1/5th of a micron necessary for the accurate identification and colocalization of subcellular domains and structures. Hence, fluorescence microscopy is routinely used to determine spatial and topological information about cells and tissues architecture, subcellular and subnuclear localization of specific gene loci, RNA molecules and proteins, from which hints into molecular functions can be inferred. Several projects at the UMR use techniques of fluorescence in situ hybridization (FISH) for the detection of specific genetic loci and RNA molecules and techniques of direct and indirect immunofluorescence for the detection of protein factors.
Use of microscopes is strictly restricted to trained users and approval of the people cited below.
Claire Francastel and Véronique Dubreuil have a long experience of these techniques and can train new users to the use of microscopes as well as image analysis.