Comparison of various sample processing methods aiming at improvement of immunogold labeling

0371068 - ÚMG 2012 GB eng A - Abstract
Sobol, Margaryta - Philimonenko, Vlada - Philimonenko, Anatoly - Hozák, Pavel
Comparison of various sample processing methods aiming at improvement of immunogold labeling.
3rd High Pressure Freezing Meeting. Londýn: UCL, London, 2011. ---.
[High Pressure Freezing Meeting /3./. 13.07.2011-15.07.2011, Londýn]
R&D Projects: GA AV ČR KAN200520704; GA MŠMT 2B06063; GA MŠMT LC545
Institutional research plan: CEZ:AV0Z50520514
Keywords : HPF * AFS * resin embedding * immunogold
Subject RIV: EB - Genetics ; Molecular Biology

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.
Permanent Link: http://hdl.handle.net/11104/0204698