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VERSION:2.0
PRODID:-//Memento EPFL//
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SUMMARY:VISUALIZATION OF NEURONAL NETWORKS IN THE MOUSE BRAIN AND MOUSE EM
 BRYOS BY ULTRAMICROSCOPY
DTSTART:20100517T161500
DTSTAMP:20260916T002722Z
UID:1c241674ff82ef2484625180e218af1b4fd3f1b86f43160a8f15be68
CATEGORIES:Conferences - Seminars
DESCRIPTION:Prof. H.U. Dodt\, Department of Bioelectronics\, Institute of 
 Solid State Electronics\, TU Vienna\, Vienna\, Austria \nIt would be very 
 helpful for the analysis of neuronal networks of the brain\, if one could 
 visualize these networks in 3 dimensions. Up to now this was only possible
  with limited resolution by sequential slicing and reconstruction of the b
 rain. This time consuming attempt is easily hampered by artifacts as shrin
 kage and distortion induced by standard histological procedures. \nTo over
 come these problems we used a microscopy based on extreme darkfield illumi
 nation with a light sheet\, once called ultramicroscopy. This microscopy a
 llows optical sectioning of whole mouse brains and was combined with an ap
 proach to clear fixed neuronal tissue: Mouse brains were made completely t
 ransparent by immersion in oil of the same refractive index as protein. By
  illuminating the brains with blue light (ë = 488 nm)\, neurons labeled w
 ith GFP were visualized by fluorescence. This way we could detect single n
 eurons in hippocampi inside whole brains. \nBy surface rendering\, the sha
 pe and position of hippocampi relative to the brain surface could be depic
 ted. In complete excised hippocampi subcellular resolution was obtained by
  3D reconstruction from several hundred optical sections. The dendritic tr
 ees of CA1 hippocampal neurons with dendrites and dendritic spines could b
 e visualized. \nMany proteins can be labeled in transgenic mice with genet
 ically encoded fluorescent markers. Using these markers our approach will 
 represent a high-throughput screening method for protein expression in 3 D
 . This expression can be monitored with µm resolution and should allow th
 e elucidation of complex neuronal networks in the brain.\nWe show that ult
 ramicroscopy also allows optical sectioning and detailed 3D reconstruction
  of whole mouse embryos by imaging autofluorescent structures. Especially 
 the circulatory system in the body and brain became apparent as blood rema
 ining in the preparation showed strong fluorescence.  Also other applicati
 ons like e.g. visualization of nerve bundles in whole embryos and visualiz
 ation of plaques in brains of mice with Alzheimers disease will be shown. 
 In general the method is well suited for high-throughput phenotype screeni
 ng of transgenic mice and thus will benefit the investigation of disease
LOCATION:AI 1-153
STATUS:CONFIRMED
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