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Optimalization of immunogold labeling on thin resin sections after cryofixation

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    0336266 - ÚMG 2010 RIV AT eng C - Conference Paper (international conference)
    Hozák, Pavel - Philimonenko, Anatoly - Philimonenko, Vlada - Sobol, Margaryta - Janáček, Jiří - Vyhnal, Aleš - Kubínová, Lucie - Janda, Pavel
    Optimalization of immunogold labeling on thin resin sections after cryofixation.
    MC 2009 - Microscopy Conference: First Joint Meeting of Dreiländertagung and Multinational Conference on Microscopy. Graz: Verlag der Technischen Universität, 2009, s. 233-234. ISBN 978-3-85125-062-6.
    [MC 2009 - Joint Meeting of Dreiländertagung and Multinational Congress on Microscopy /9./. Graz (AT), 30.08.2009-04.09.2009]
    R&D Projects: GA AV ČR KAN200520704; GA MŠk LC545; GA MŠk 2B06063; GA ČR GD204/09/H084
    Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z50110509; CEZ:AV0Z40400503
    Keywords : cell ultrastructure * high pressure freezing * freeze substitutions * resins * immunocytochemistry
    Subject RIV: EB - Genetics ; Molecular Biology

    Various 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.
    Permanent Link: http://hdl.handle.net/11104/0180531

     
     
Number of the records: 1