Počet záznamů: 1

Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding

  1. 1.
    0355818 - UMG-J 2011 RIV DE eng J - Článek v odborném periodiku
    Sobol, Margaryta - Philimonenko, Vlada - Hozák, Pavel
    Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding.
    Histochemistry and Cell Biology. Roč. 134, č. 6 (2010), s. 631-641 ISSN 0948-6143
    Grant CEP: GA AV ČR KAN200520704; GA MŠk 2B06063; GA MŠk LC545
    Výzkumný záměr: CEZ:AV0Z50520514
    Klíčová slova: cryofixation * automated freeze-substitution * LR White
    Kód oboru RIV: EB - Genetika a molekulární biologie
    Impakt faktor: 4.727, rok: 2010

    In this study we present an optimized method of high-pressure freezing and automated freeze-substitution of cultured human cells, followed by LR White embedding, for subsequent immunolabeling. Also, the influence of various conditions of the freeze-substitution procedures such as temperature, duration, and additives in the substitution medium on the preservation of cryo-immobilized cells was analyzed. The recommended approach combines (1) automated freeze-substitution for high reproducibility and minimizing human-derived errors; (2) minimal addition of contrasting and fixing agents; (3) easy-to-use LR White resin for embedment; (4) good preservation of nuclei and nucleoli which are usually the most difficult structures to effectively vitrify and saturate in a resin; and (5) preservation of antigens for sensitive immunogold labeling.
    Trvalý link: http://hdl.handle.net/11104/0194499