Improvements in immunostaining samples embedded in methacrylate: localization of microtubules and other antigens throughout developing organs in plants of diverse taxa.

摘要: Microtubules are important in plant growth and development. Localizing microtubules sectioned material is advantageous because it allows any tissue of interest to be studied permits the positional relations cells within organ known. We describe here a method that uses semi-thin (0.5–2 μm) sections embedded butyl-methylmethacrylate, which 10 mM dithiothreitol was added. After removing embedding using indirect immunofluorescence staining, we obtain clear images microtubules, actin microfilaments, callose pulse-fed bromodeoxyuridine. This works on root tissues Arabidopsis thaliana(L.) Heynh, Pinus radiataD. Don, Zamia furfuraceaAit., Azolla pinnataR. Br. sporophytic Funaria hygrometricaHedw. In general, most organs successfully stained. Using this method, find interphase meristematic all these species have not only usual cortical array but also throughout their cytoplasm. The presence calcium chelator ethylene glycol-bis(β-aminoethyl ether)N,N,N′,N′-tetraacetic acid EGTA fixation buffers led some damage, did enhance preservation microtubules. common assumption EGTA-containing stabilize during appears unwarranted.