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Comparison of various sample processing methods aiming at improvement of immunogold labeling
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SYSNO ASEP 0371068 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 Comparison of various sample processing methods aiming at improvement of immunogold labeling Author(s) Sobol, Margaryta (UMG-J) RID
Philimonenko, Vlada (UMG-J)
Philimonenko, Anatoly (UMG-J)
Hozák, Pavel (UMG-J) RID, ORCIDSource Title 3rd High Pressure Freezing Meeting. - Londýn : UCL, London, 2011
---Number of pages 1 s. Action High Pressure Freezing Meeting /3./ Event date 13.07.2011-15.07.2011 VEvent location Londýn Country GB - United Kingdom Event type EUR Language eng - English Country GB - United Kingdom Keywords HPF ; AFS ; resin embedding ; immunogold Subject RIV EB - Genetics ; Molecular Biology R&D Projects KAN200520704 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) 2B06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LC545 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) CEZ AV0Z50520514 - UMG-J (2005-2011) Annotation We present our experiments aiming to improve the preservation of fine ultrastructural details and antigenicity of biological samples using high-pressure freezing (HPF) followed by automated freeze-substitution (FS) and resin embedment for the subsequent immunogold labeling. We assessed different non-penetrating cryoprotectants such as fetal bovine serum (FBS), low-temperature gelling agarose, and bovine serum albumin (BSA) in various mixtures for HPF to minimize the appearance of segregation patterns and attain a good preservation of an ultrastructure of cellular components, especially nuclei, nucleoli, and mitotic chromatin. We also estimated the ability of various embedding resins LR White, LR Gold, Lowicryl K4M, and Lowicryl HM20 for sensitive revealing the antigens on thin sections by immunogold labeling. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2012
Number of the records: 1