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How to improve preparation of samples for immunogold labeling on ultrathin sections?
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SYSNO ASEP 0370859 Document Type A - Abstract R&D Document Type The record was not marked in the RIV R&D Document Type Není vybrán druh dokumentu Title How to improve preparation of samples for immunogold labeling on ultrathin sections? Author(s) Sobol, Margaryta (UMG-J) RID
Philimonenko, Vlada (UMG-J)
Philimonenko, Anatoly (UMG-J)
Janda, Pavel (UFCH-W) RID, ORCID
Nebesářová, Jana (BC-A) RID, ORCID
Hozák, Pavel (UMG-J) RID, ORCIDSource Title Mikroskopie 2011. - Praha : Československá mikroskopická společnost, 2011
S. 31-31Number of pages 1 s. Action Mikroskopie 2011 Event date 17.02.2011-18.02.2011 VEvent location Nové Město na Moravě Country CZ - Czech Republic Event type EUR Language eng - English Country CZ - Czech Republic Keywords HPF/FS ; 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) CEZ AV0Z50520514 - UMG-J (2005-2011) Annotation Method for preservation of ultrastructure and antigenicity of human cultured interphase and mitotic cells allowing subsequent immunogold labeling on thin sections was presented, combining HPF and FS followed by resin embedding with some essential recommendations. Various HPF cryoprotectants to achieve an improved cell ultrastructure, the cryoprotection method of choice were discussed. Effective FS procedure of cryofixed cells using glutaraldehyde and water added to acetone minimize the sample structural distortions. FS protocol followed by LR White embedding favors preservation of both fine morphology and contrast in interphase cells, but also in mitotic cells. Detailed protocol was presented, with comparison of several embedding resins for their ability to expose antigens on ultrathin sections for immunolabeling. LR White resin in combination with HPF/FS for fine ultrastructural immunocytochemistry gives good preservation of both fine ultrastructure and immunolabeling intensity. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2012
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