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

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	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. 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. (en)
Title	Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding (en)
skos:prefLabel	Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding (en)
skos:notation	RIV/68378050:_____/10:00355818!RIV11-MSM-68378050
http://linked.open...avai/riv/aktivita	P Z
http://linked.open...avai/riv/aktivity	P(2B06063), P(KAN200520704), P(LC545), Z(AV0Z50520514)
http://linked.open...iv/cisloPeriodika	6
http://linked.open...vai/riv/dodaniDat	2011
http://linked.open...aciTvurceVysledku	Sobol, Margaryta Hozák, Pavel Philimonenko, Vlada
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Ústav molekulární genetiky AV ČR, v. v. i.
http://linked.open...dnocenehoVysledku	251476
http://linked.open...ai/riv/idVysledku	RIV/68378050:_____/10:00355818
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	cryofixation; automated freeze-substitution; LR White (en)
http://linked.open.../riv/klicoveSlovo	LR White automated freeze-substitution cryofixation
http://linked.open...odStatuVydavatele	DE - Spolková republika Německo
http://linked.open...ontrolniKodProRIV	[A5450DE9D561]
http://linked.open...i/riv/nazevZdroje	Histochemistry and Cell Biology
http://linked.open...in/vavai/riv/obor	EB
http://linked.open...ichTvurcuVysledku	3 (xsd:int)
http://linked.open...cetTvurcuVysledku	3 (xsd:int)
http://linked.open...vavai/riv/projekt	Structure and irregularities of the nucleoskeleton: identification and diagnostics. Functional Organisation of the Cell New nanoparticles for ultrastructural diagnostics
http://linked.open...UplatneniVysledku	2010
http://linked.open...v/svazekPeriodika	134
http://linked.open...iv/tvurceVysledku	Hozák, Pavel Sobol, Margaryta Philimonenko, Vlada
http://linked.open...ain/vavai/riv/wos	000284645000010
http://linked.open...n/vavai/riv/zamer	Molekulárně genetické a buněčné základy klíčových biologických procesů: genová exprese, onkogeneze, replikace virů, imunita a vývoj organismů
issn	0948-6143
number of pages	11 (xsd:int)
is http://linked.open...avai/riv/vysledek of	Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding

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