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Fluorescent in situ visualization of sterols in Arabidopsis roots
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Umeå Plant Science Centre (UPSC).
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik. Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Umeå Plant Science Centre (UPSC).ORCID-id: 0000-0001-7571-0670
2011 (engelsk)Inngår i: Nature Protocols, ISSN 1754-2189, E-ISSN 1750-2799, Vol. 6, nr 4, s. 446-456Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Sterols are eukaryotic membrane components with crucial roles in diverse cellular processes. Elucidation of sterol function relies on development of tools for in situ sterol visualization. Here we describe protocols for in situ sterol localization in Arabidopsis thaliana root cells, using filipin as a specific probe for detection of fluorescent filipin-sterol complexes. Currently, filipin is the only established tool for sterol visualization in plants. Filipin labeling can be performed on aldehyde-fixed samples, largely preserving fluorescent proteins and being compatible with immunocytochemistry. Filipin can also be applied for probing live cells, taking into account the fact that it inhibits sterol-dependent endocytosis. The experimental procedures described are designed for fluorescence detection by confocal laser-scanning microscopy with excitation of filipin-sterol complexes at 364 nm. The protocols require 1 d for sterol covisualization with fluorescent proteins in fixed or live roots and 2 d for immunocytochemistry on whole-mount roots.

sted, utgiver, år, opplag, sider
2011. Vol. 6, nr 4, s. 446-456
Emneord [en]
Imaging, Model organisms, Plant biology
HSV kategori
Identifikatorer
URN: urn:nbn:se:umu:diva-41115DOI: 10.1038/nprot.2011.323PubMedID: 21412273OAI: oai:DiVA.org:umu-41115DiVA, id: diva2:404725
Tilgjengelig fra: 2011-03-18 Laget: 2011-03-18 Sist oppdatert: 2018-03-15bibliografisk kontrollert

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