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Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding
M. Sobol, VV. Philimonenko, P. Hozák
Jazyk angličtina Země Německo
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem
NLK
ProQuest Central
od 1997-01-01 do Před 1 rokem
Medline Complete (EBSCOhost)
od 2000-01-01 do Před 1 rokem
Nursing & Allied Health Database (ProQuest)
od 1997-01-01 do Před 1 rokem
Health & Medicine (ProQuest)
od 1997-01-01 do Před 1 rokem
Public Health Database (ProQuest)
od 1997-01-01 do Před 1 rokem
- MeSH
- akrylové pryskyřice diagnostické užití MeSH
- elektronová mikroskopie MeSH
- HeLa buňky ultrastruktura MeSH
- histologické techniky metody MeSH
- imunohistochemie metody MeSH
- lidé MeSH
- mrazová substituce metody MeSH
- ochrana biologická metody MeSH
- tlak MeSH
- zalévání tkání metody MeSH
- zmrazování MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
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.
Citace poskytuje Crossref.org
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- $a Sobol, Margarita $u Department of Biology of the Cell Nucleus, Institute of Molecular Genetics of the Academy of Sciences of the Czech Republic, Prague, Czech Republic
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- $a Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding / $c M. Sobol, VV. Philimonenko, P. Hozák
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- $a 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.
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- $a Philimonenko, Vlada V. $u Institute of Experimental Medicine, Department of Cell Ultrastructure and Molecular Biology, Academy of Sciences of the Czech Republic, Vídenska 1083, 142 20 Prague 4 - Krc, Czech Republic
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