Immunofluorescence detection of F-actin on low melting point wax sections from plant tissues.
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-hydroxysuccinimide 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.
Vitha
S
S
Institute of Plant Molecular Biology, Academy of Sciences of Czech Republic, Ceské Budĕjovice, Czech Republic.
Baluska
F
F
Mews
M
M
Volkmann
D
D
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000648:ultrastructure*; D000199:Actins / Q000032:analysis*; D019084:Fluorescent Antibody Technique, Indirect; D018517:Plant Roots / Q000737:chemistry*; D018547:Plant Stems / Q000737:chemistry*; D013696:Temperature; D016610:Tissue Embedding; D016707:Tissue Fixation; D003313:Zea mays
D000199:Actins