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Journal of Histochemistry and Cytochemistry, Vol. 45, 89-96, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

Immunofluorescence Detection of F-actin on Low Melting Point Wax Sections from Plant Tissues

Stanislav Vithaa, Frantisek Baluskab,c, Miriam Mewsc, and Dieter Volkmannc
a Institute of Plant Molecular Biology, Academy of Sciences of Czech Republic, Ceské Budejovice, Czech Republic
b Institute of Botany, Slovak Academy of Sciences, Bratislava, Slovakia
c Botanisches Institut der Universität Bonn, Bonn, Germany

Correspondence to: Stanislav Vitha, Plant Biology, Ohio State U., 1735 Neil Ave., Columbus, OH 43210.

We developed a simple and reliable technique for immunofluorescence detection of F-actin on microtome sections of plant tissues. For the first time, large numbers of plant cells from various tissues that pass through their developmental stages could be consistently visualized on one section from plant organs. n-Maleimidobenzoic acid N-hydroxy-succinimide ester-pretreated and formalin-fixed segments of plant roots and shoots were embedded in low melting point ester wax at 37C and sectioned on a microtome. After dewaxing and rehydration, microfilaments were visualized by indirect immunofluorescence technique with a monoclonal anti-actin antibody. The technique has been successfully used for visualization of tissue- and development-specific F-actin arrays in cells of Zea mays and Lepidium sativum root tips and of maize stem nodes. (J Histochem Cytochem 45:89-95, 1997)

Key Words: Fixation, Actin, Cytoskeleton, Immunofluorescence, Maize root


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