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@ARTICLE{Marx:19855,
      author       = {Marx, M. and Günter, R.H. and Hucko, W. and Radnikow, G.
                      and Feldmeyer, D.},
      title        = {{I}mproved biocytin labeling and neuronal 3{D}
                      reconstruction},
      journal      = {Nature protocols},
      volume       = {7},
      issn         = {1754-2189},
      address      = {Basingstoke},
      publisher    = {Nature Publishing Group},
      reportid     = {PreJuSER-19855},
      pages        = {394 - 407},
      year         = {2012},
      note         = {This work was supported by the Deutsche
                      Forschungsgemeinschaft (DFG; Research Group BaCoFun), the
                      Helmholtz Association and the Helmholtz Alliance for Systems
                      Biology. We thank A. Rollenhagen for help with the EM
                      protocol and T. Abel for critically reading the manuscript.},
      abstract     = {In this report, we describe a reliable protocol for
                      biocytin labeling of neuronal tissue and diaminobenzidine
                      (DAB)-based processing of brain slices. We describe how to
                      embed tissues in different media and how to subsequently
                      histochemically label the tissues for light or electron
                      microscopic examination. We provide a detailed dehydration
                      and embedding protocol using Eukitt that avoids the common
                      problem of tissue distortion and therefore prevents fading
                      of cytoarchitectural features (in particular, lamination) of
                      brain tissue; as a result, additional labeling methods (such
                      as cytochrome oxidase staining) become unnecessary. In
                      addition, we provide correction factors for tissue shrinkage
                      in all spatial dimensions so that a realistic neuronal
                      morphology can be obtained from slice preparations. Such
                      corrections were hitherto difficult to calculate because
                      embedding in viscous media resulted in highly nonlinear
                      tissue deformation. Fixation, immunocytochemistry and
                      embedding procedures for light microscopy (LM) can be
                      completed within 42-48 h. Subsequent reconstructions and
                      morphological analyses take an additional 24 h or more.},
      keywords     = {Animals / Brain: cytology / Brain: ultrastructure /
                      Imaging, Three-Dimensional: methods / Lysine: analogs $\&$
                      derivatives / Mice / Microtomy: methods / Neurons:
                      ultrastructure / Osmium Tetroxide / Rats / Staining and
                      Labeling: methods / Osmium Tetroxide (NLM Chemicals) /
                      Lysine (NLM Chemicals) / biocytin (NLM Chemicals) / J
                      (WoSType)},
      cin          = {INM-2},
      ddc          = {610},
      cid          = {I:(DE-Juel1)INM-2-20090406},
      pnm          = {Funktion und Dysfunktion des Nervensystems (FUEK409) /
                      89571 - Connectivity and Activity (POF2-89571)},
      pid          = {G:(DE-Juel1)FUEK409 / G:(DE-HGF)POF2-89571},
      shelfmark    = {Biochemical Research Methods},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:22301777},
      UT           = {WOS:000300402300015},
      doi          = {10.1038/nprot.2011.449},
      url          = {https://juser.fz-juelich.de/record/19855},
}