Ratiometric Fluorescence Live Imaging Analysis of Membrane Lipid Order in Arabidopsis Mitotic Cells Using a Lipid Order-Sensitive Probe.
Methods Mol Biol
; 1370: 227-39, 2016.
Article
em En
| MEDLINE
| ID: mdl-26659966
ABSTRACT
Eukaryotic cells contain membranes exhibiting different levels of lipid order mostly related to their relative amount of sterol-rich domains, thought to mediate temporal and spatial organization of cellular processes. We previously provided evidence in Arabidopsis thaliana that sterols are crucial for execution of cytokinesis, the last stage of cell division. Recently, we used di-4-ANEPPDHQ, a fluorescent probe sensitive to order of lipid phases, to quantify the level of membrane order of the cell plate, the membrane structure separating daughter cells during somatic cytokinesis of higher plant cells. By employing quantitative, ratiometric fluorescence microscopy for mapping localized lipid order levels, we revealed that the Arabidopsis cell plate represents a high-lipid-order domain of the plasma membrane. Here, we describe step-by-step protocols and troubleshooting for ratiometric live imaging procedures employing the di-4-ANEPPDHQ fluorescent probe for quantification of membrane lipid order during plant cell division in suspension cell cultures and roots of Arabidopsis thaliana.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Compostos de Piridínio
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Arabidopsis
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Corantes Fluorescentes
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Lipídeos de Membrana
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Microscopia de Fluorescência
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Mitose
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article