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Confocal Microscopy
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Confocal microscopy extends the capability of conventional fluorescence microscopy by enabling the investigator to visualize a thin optical plane within a sample. Out of focus portions of the sample are minimized in the image produced. Optical sections obtained confocally are superior to those obtained by mechanical sectioning, because consecutive sections remain in precise register and can be used to produce an accurate three dimensional image or model of the original sample. In 2007 a new Leica TCS SP5 inverted confocal microscope was added to the two Zeiss 510 systems (one upright the other inverted) that are available for use by members of the MRDDRC. These three systems incorporate many refinements and advances that extend their usefulness for biological research. For example, the x,y,z stages allow automated collection of high resolution montage images of samples far larger than the field of view of a particular lens. Improved automation of many collection and measurement functions both simplify use and improve mechanical precision of the systems. Time lapse and bleaching capabilities allow one to use recent methods such as FRAP to study motility of molecules within membranes and FRET to determine the extent of separation of molecules in the nanometer range. The new Leica system adds additional capabilities for live cell imaging through the use of a high speed resonance scanner, greater flexibility of usable fluorochromes through the addition of a UV laser and a forth detector and through a more precise means to select emission wavelengths. Each microscope has an independent file server and an imaging workstation with software for preparation of publication quality figures. The Confocal Microscopy Core provides training and ongoing support for MRDDRC users of core equipment for a reduced fee.
Use of these systems by MRDDRC investigators has facilitated discovery and elucidation of genes involved in human disorders such as Rett and SCA1; functional analyses of genes including Math1, Mecp2, Ataxin, Gfi-1, shar-pei, VAP-33A, endophilin, crumbs, hrs, CSP, atonal, syntaxin, senseless, neurexin, ROP, discs lost, skittles, gutfeeling, synaptobrevin, synaptotagmin, apterous, Dpp and protein structure/function studies of potassium channel domains.
For more information, please contact Richard Atkinson.
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