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Simultaneous detection of multiple targets for ultrastructural immunocytochemistry
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SYSNO ASEP 0434293 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Simultaneous detection of multiple targets for ultrastructural immunocytochemistry Author(s) Philimonenko, Vlada (UMG-J)
Filimonenko, Anatolij (UMG-J)
Šloufová, I. (CZ)
Hrubý, Martin (UMCH-V) RID, ORCID
Novotný, F. (CZ)
Halbhuber, Z. (CZ)
Krivjanská, M. (CZ)
Nebesářová, Jana (BC-A) RID, ORCID
Šlouf, Miroslav (UMCH-V) RID, ORCID
Hozák, Pavel (UMG-J) RID, ORCIDSource Title Histochemistry and Cell Biology. - : Springer - ISSN 0948-6143
Roč. 141, č. 3 (2014), s. 229-239Number of pages 11 s. Language eng - English Country DE - Germany Keywords Immunolabeling ; Metal nanoparticles ; Electron microscopy ; Cell nucleus ; Ultrastructure ; Phosphatidylinositol-4,5-Bisphosphate (PIP2) Subject RIV EB - Genetics ; Molecular Biology R&D Projects KAN200520704 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) TE01020118 GA TA ČR - Technology Agency of the Czech Republic (TA ČR) GAP305/11/2232 GA ČR - Czech Science Foundation (CSF) LD12063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UMCH-V - RVO:61389013 ; BC-A - RVO:60077344 ; UMG-J - RVO:68378050 CEZ AV0Z50520514 - UMG-J (2005-2011) UT WOS 000332082300001 DOI 10.1007/s00418-013-1178-6 Annotation Simultaneous detection of biological molecules by means of indirect immunolabeling provides valuable information about their localization in cellular compartments and their possible interactions in macromolecular complexes. While fluorescent microscopy allows for simultaneous detection of multiple antigens, the sensitive electron microscopy immunodetection is limited to only two antigens. In order to overcome this limitation, we prepared a set of novel, shape-coded metal nanoparticles readily discernible in transmission electron microscopy which can be conjugated to antibodies or other bioreactive molecules. With the use of novel nanoparticles, various combinations with commercial gold nanoparticles can be made to obtain a set for simultaneous labeling. For the first time in ultrastructural histochemistry, up to five molecular targets can be identified simultaneously. We demonstrate the usefulness of the method by mapping of the localization of nuclear lipid phosphatidylinositol-4,5-bisphosphate together with four other molecules crucial for genome function, which proves its suitability for a wide range of biomedical applications. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2015
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