How to improve preparation of samples for immunogold labeling on ultrathin sections?

0370859 - ÚMG 2012 CZ eng A - Abstrakt
Sobol, Margaryta - Philimonenko, Vlada - Philimonenko, Anatoly - Janda, Pavel - Nebesářová, Jana - Hozák, Pavel
How to improve preparation of samples for immunogold labeling on ultrathin sections?
Mikroskopie 2011. Praha: Československá mikroskopická společnost, 2011. s. 31-31.
[Mikroskopie 2011. 17.02.2011-18.02.2011, Nové Město na Moravě]
Grant CEP: GA AV ČR KAN200520704; GA MŠMT LC545; GA MŠMT 2B06063
Výzkumný záměr: CEZ:AV0Z50520514
Klíčová slova: HPF/FS * resins * immunocytochemistry
Kód oboru RIV: EB - Genetika a molekulární biologie

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.
Trvalý link: http://hdl.handle.net/11104/0204541