caa a22 4500 467901104 CHVBK 20180406152818.0 cr unu---uuuuu 170328e20060801xx s 000 0 eng 10.1007/s00299-005-0089-y doi (NATIONALLICENCE)springer-10.1007/s00299-005-0089-y Visualisation of microtubules and actin filaments in fixed BY-2 suspension cells using an optimised whole mount immunolabelling protocol [Elektronische Daten] [Magdalena Szechyńska-Hebda, Maria Wędzony, Ewa Dubas, Henk Kieft, André van Lammeren] Excellent visualisation of microtubules and actin filaments was obtained in fixed tobacco BY-2 suspension cells after optimising a protocol for whole mount immunolabelling. The procedure is based on modification of fixation, cell wall digestion, dimethyl sulfoxide (DMSO) treatment, post fixation, and blocking. The most critical aspects of successful preservation and visualization of cytoskeletal elements appeared to be: a two-step fixation with paraformaldehyde and glutaraldehyde before enzymatic cell wall digestion and a post fixation with aldehydes thereafter. The method allows the improved visualization of the organisation of the microtubular and actin filament arrays during the successive stages of cell division and at interphase. Although we present the application of our protocols for cytoskeleton labelling, the excellent results show the potential of using this method for the analysis of various proteins and molecules in plant cells. Springer-Verlag, 2005 Actin filaments nationallicence Cell division nationallicence Cytoskeleton nationallicence Immunocytochemistry nationallicence Microtubules nationallicence Tobacco BY-2 cells nationallicence AF : Actin filament nationallicence BSA : Bovine serum albumin nationallicence BY-2 cells : Tobacco Bright Yellow-2 cells nationallicence DMSO : Dimethyl sulfoxide nationallicence GFP : Green fluorescent protein nationallicence MBD : Microtubule binding domain of the microtubule associated protein 4 nationallicence MBS ester : 3-Maleimidobenzoyl-N-hydroxysuccinimide ester nationallicence MSB : Microtubule stabilizing buffer nationallicence MT : Microtubule nationallicence PFA : Paraformaldehyde nationallicence Szechyńska-Hebda Magdalena Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut Wędzony Maria Institute of Plant Physiology Polish Academy of Sciences, ul. Niezapominajek 21, 30-239, Kraków, Poland aut Dubas Ewa Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut Kieft Henk Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut van Lammeren André Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut Plant Cell Reports Springer-Verlag 25/8(2006-08-01), 758-766 0721-7714 25:8<758 2006 25 299 https://doi.org/10.1007/s00299-005-0089-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0089-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Szechyńska-Hebda Magdalena Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Wędzony Maria Institute of Plant Physiology Polish Academy of Sciences, ul. Niezapominajek 21, 30-239, Kraków, Poland aut NATIONALLICENCE 700 1- Dubas Ewa Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- Kieft Henk Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut NATIONALLICENCE 700 1- van Lammeren André Laboratory for Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/8(2006-08-01), 758-766 0721-7714 25:8<758 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer