000019855 001__ 19855 000019855 005__ 20210129210736.0 000019855 0247_ $$2pmid$$apmid:22301777 000019855 0247_ $$2DOI$$a10.1038/nprot.2011.449 000019855 0247_ $$2WOS$$aWOS:000300402300015 000019855 0247_ $$2altmetric$$aaltmetric:896846 000019855 037__ $$aPreJuSER-19855 000019855 041__ $$aeng 000019855 082__ $$a610 000019855 084__ $$2WoS$$aBiochemical Research Methods 000019855 1001_ $$0P:(DE-Juel1)VDB6549$$aMarx, M.$$b0$$uFZJ 000019855 245__ $$aImproved biocytin labeling and neuronal 3D reconstruction 000019855 260__ $$aBasingstoke$$bNature Publishing Group$$c2012 000019855 300__ $$a394 - 407 000019855 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article 000019855 3367_ $$2DataCite$$aOutput Types/Journal article 000019855 3367_ $$00$$2EndNote$$aJournal Article 000019855 3367_ $$2BibTeX$$aARTICLE 000019855 3367_ $$2ORCID$$aJOURNAL_ARTICLE 000019855 3367_ $$2DRIVER$$aarticle 000019855 440_0 $$024802$$aNature Protocols$$v7 000019855 500__ $$aThis 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. 000019855 520__ $$aIn this report, we describe a reliable protocol for biocytin labeling of neuronal tissue and diaminobenzidine (DAB)-based processing of brain slices. We describe how to embed tissues in different media and how to subsequently histochemically label the tissues for light or electron microscopic examination. We provide a detailed dehydration and embedding protocol using Eukitt that avoids the common problem of tissue distortion and therefore prevents fading of cytoarchitectural features (in particular, lamination) of brain tissue; as a result, additional labeling methods (such as cytochrome oxidase staining) become unnecessary. In addition, we provide correction factors for tissue shrinkage in all spatial dimensions so that a realistic neuronal morphology can be obtained from slice preparations. Such corrections were hitherto difficult to calculate because embedding in viscous media resulted in highly nonlinear tissue deformation. Fixation, immunocytochemistry and embedding procedures for light microscopy (LM) can be completed within 42-48 h. Subsequent reconstructions and morphological analyses take an additional 24 h or more. 000019855 536__ $$0G:(DE-Juel1)FUEK409$$2G:(DE-HGF)$$aFunktion und Dysfunktion des Nervensystems (FUEK409)$$cFUEK409$$x0 000019855 536__ $$0G:(DE-HGF)POF2-89571$$a89571 - Connectivity and Activity (POF2-89571)$$cPOF2-89571$$fPOF II T$$x1 000019855 588__ $$aDataset connected to Web of Science, Pubmed 000019855 650_2 $$2MeSH$$aAnimals 000019855 650_2 $$2MeSH$$aBrain: cytology 000019855 650_2 $$2MeSH$$aBrain: ultrastructure 000019855 650_2 $$2MeSH$$aImaging, Three-Dimensional: methods 000019855 650_2 $$2MeSH$$aLysine: analogs & derivatives 000019855 650_2 $$2MeSH$$aMice 000019855 650_2 $$2MeSH$$aMicrotomy: methods 000019855 650_2 $$2MeSH$$aNeurons: ultrastructure 000019855 650_2 $$2MeSH$$aOsmium Tetroxide 000019855 650_2 $$2MeSH$$aRats 000019855 650_2 $$2MeSH$$aStaining and Labeling: methods 000019855 650_7 $$020816-12-0$$2NLM Chemicals$$aOsmium Tetroxide 000019855 650_7 $$056-87-1$$2NLM Chemicals$$aLysine 000019855 650_7 $$0576-19-2$$2NLM Chemicals$$abiocytin 000019855 650_7 $$2WoSType$$aJ 000019855 7001_ $$0P:(DE-Juel1)VDB98246$$aGünter, R.H.$$b1$$uFZJ 000019855 7001_ $$0P:(DE-Juel1)VDB104574$$aHucko, W.$$b2$$uFZJ 000019855 7001_ $$0P:(DE-Juel1)VDB36676$$aRadnikow, G.$$b3$$uFZJ 000019855 7001_ $$0P:(DE-Juel1)131680$$aFeldmeyer, D.$$b4$$uFZJ 000019855 773__ $$0PERI:(DE-600)2244966-8$$a10.1038/nprot.2011.449$$gVol. 7, p. 394 - 407$$p394 - 407$$q7<394 - 407$$tNature protocols$$v7$$x1754-2189$$y2012 000019855 8567_ $$uhttp://dx.doi.org/10.1038/nprot.2011.449 000019855 909CO $$ooai:juser.fz-juelich.de:19855$$pVDB 000019855 9132_ $$0G:(DE-HGF)POF3-571$$1G:(DE-HGF)POF3-570$$2G:(DE-HGF)POF3-500$$aDE-HGF$$bKey Technologies$$lDecoding the Human Brain$$vConnectivity and Activity$$x0 000019855 9131_ $$0G:(DE-HGF)POF2-89571$$1G:(DE-HGF)POF3-890$$2G:(DE-HGF)POF3-800$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bProgrammungebundene Forschung$$lohne Programm$$vConnectivity and Activity$$x1 000019855 9141_ $$y2012 000019855 915__ $$0StatID:(DE-HGF)0040$$2StatID$$aPeer review unknown 000019855 915__ $$0StatID:(DE-HGF)0111$$2StatID$$aWoS$$bScience Citation Index Expanded 000019855 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection 000019855 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bThomson Reuters Master Journal List 000019855 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS 000019855 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline 000019855 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database 000019855 915__ $$0StatID:(DE-HGF)0420$$2StatID$$aNationallizenz 000019855 915__ $$0StatID:(DE-HGF)1030$$2StatID$$aDBCoverage$$bCurrent Contents - Life Sciences 000019855 9201_ $$0I:(DE-Juel1)INM-2-20090406$$gINM$$kINM-2$$lMolekulare Organisation des Gehirns$$x0 000019855 970__ $$aVDB:(DE-Juel1)134869 000019855 980__ $$aVDB 000019855 980__ $$aConvertedRecord 000019855 980__ $$ajournal 000019855 980__ $$aI:(DE-Juel1)INM-2-20090406 000019855 980__ $$aUNRESTRICTED