Using FM Dyes to Study Endomembranes and Their Dynamics in Plants and Cell Suspensions
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Arabidopsis thaliana root tip, BY-2, Endocytosis, Endomembranes, FM styryl dyes, Microscopy, Plasma membrane,
- MeSH
- Arabidopsis cytology ultrastructure MeSH
- Cell Membrane ultrastructure MeSH
- Endocytosis * MeSH
- Fluorescent Dyes analysis MeSH
- Microscopy, Fluorescence methods MeSH
- Plant Roots cytology ultrastructure MeSH
- Optical Imaging methods MeSH
- Nicotiana cytology ultrastructure MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Fluorescent Dyes MeSH
FM (Fei-Mao) styryl dyes are commonly used for the fluorescence imaging of plasma membrane (PM) and endocytosis in vivo. Thanks to their amphiphilic character, these dyes are incorporated in the outer leaflet of the PM lipid bilayer and emit fluorescence in its hydrophobic environment. The endocytic pathway of FM dye uptake starts with rapid PM staining and continues in PM invaginations and membrane vesicles during endocytosis, followed by staining of trans-Golgi network (TGN) and ending in tonoplast (vacuolar membrane). FM dyes do not stain endoplasmic reticulum and nuclear membrane. The time-lapse fluorescence microscopy could track endocytic vesicles and characterize the rate of endocytosis in vivo. On the other hand, fixable FM dyes (FX) can be used for the visualization of particular steps in the FM dye uptake in situ. Staining with FM dyes and subsequent microscopic observations could be performed on both tissue and cellular level. Here, we describe simple procedures for the effective FM dye staining and destaining in root tip of Arabidopsis thaliana seedlings and suspension-cultured tobacco cells.
Department of Experimental Plant Biology Faculty of Science Charles University Prague Czech Republic
Institute of Experimental Botany of the Czech Academy of Sciences Prague Czech Republic
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