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Optimalization of immunogold labeling on thin resin sections after cryofixation

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    SYSNO ASEP0336266
    Document TypeC - Proceedings Paper (int. conf.)
    R&D Document TypeConference Paper
    TitleOptimalization of immunogold labeling on thin resin sections after cryofixation
    Author(s) Hozák, Pavel (UMG-J) RID
    Philimonenko, Anatoly (UMG-J)
    Philimonenko, Vlada (UMG-J)
    Sobol, Margaryta (UMG-J) RID
    Janáček, Jiří (FGU-C) RID, ORCID
    Vyhnal, Aleš (FGU-C)
    Kubínová, Lucie (FGU-C) RID, ORCID
    Janda, Pavel (UFCH-W) RID, ORCID
    Number of authors8
    Source TitleMC 2009 - Microscopy Conference: First Joint Meeting of Dreiländertagung and Multinational Conference on Microscopy. - Graz : Verlag der Technischen Universität, 2009 - ISBN 978-3-85125-062-6
    Pagess. 233-234
    Number of pages2 s.
    ActionMC 2009 - Joint Meeting of Dreiländertagung and Multinational Congress on Microscopy /9./
    Event date30.08.2009-04.09.2009
    VEvent locationGraz
    CountryAT - Austria
    Event typeWRD
    Languageeng - English
    CountryAT - Austria
    Keywordscell ultrastructure ; high pressure freezing ; freeze substitutions ; resins ; immunocytochemistry
    Subject RIVEB - Genetics ; Molecular Biology
    R&D ProjectsKAN200520704 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
    LC545 GA MŠk - Ministry of Education, Youth and Sports (MEYS)
    2B06063 GA MŠk - Ministry of Education, Youth and Sports (MEYS)
    GD204/09/H084 GA ČR - Czech Science Foundation (CSF)
    CEZAV0Z50520514 - UMG-J (2005-2011)
    AV0Z50110509 - FGU-C (2005-2011)
    AV0Z40400503 - UFCH-W (2005-2011)
    AnnotationVarious resins (Lowicryl, LR White, LR Gold, Unicryl) used for embedding were compared for their ability to reveal antigens for immunogold labelling on their surface. Their surfaces after cutting with a diamond knife were observed using atomic force microscopy, and results statistically analyzed. The LR White resin in combination with HPF/FS was shown to be able to be successfully used for fine ultrastructural immunocytochemistry. Methods were presented allowing detection of clustering or colocalization of antigens/gold particles. A complete set of tools is now available for reliable detection of gold particles in EM images, their statistical evaluation and production of convenient outputs for publication of results. The plug-ins are useful addition to image analysis software accompanying modern CCD cameras for electron microscopes.
    WorkplaceInstitute of Molecular Genetics
    ContactGabriela Marešová, maresova@img.cas.cz, Tel.: 241 063 217
    Year of Publishing2010
Number of the records: 1