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Optimalization of immunogold labeling on thin resin sections after cryofixation
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SYSNO ASEP 0336266 Document Type C - Proceedings Paper (int. conf.) R&D Document Type Conference Paper Title Optimalization of immunogold labeling on thin resin sections after cryofixation Author(s) Hozák, Pavel (UMG-J) RID, ORCID
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, ORCIDNumber of authors 8 Source Title MC 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 Pages s. 233-234 Number of pages 2 s. Action MC 2009 - Joint Meeting of Dreiländertagung and Multinational Congress on Microscopy /9./ Event date 30.08.2009-04.09.2009 VEvent location Graz Country AT - Austria Event type WRD Language eng - English Country AT - Austria Keywords cell ultrastructure ; high pressure freezing ; freeze substitutions ; resins ; immunocytochemistry Subject RIV EB - Genetics ; Molecular Biology R&D Projects KAN200520704 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) LC545 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) 2B06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GD204/09/H084 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z50520514 - UMG-J (2005-2011) AV0Z50110509 - FGU-C (2005-2011) AV0Z40400503 - UFCH-W (2005-2011) Annotation 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. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2010
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