Imaging Plant Nuclei and Membrane-Associated Cytoskeleton by Field Emission Scanning Electron Microscopy - Plant Cell Morphogenesis: Methods and Protocols - page 169

Fig. 2 Tobacco BY-2 cytoplasmic membrane exposed from cytoplasmic side and
viewed by feiSEM. ( a ) Low magnifi cation image. ( b ) and ( c ) Details of the micro-
tubule array attached to the plasma membrane. Arrows indicate the microtu-
bules, black arrowheads show microtubule associated proteins, and white
arrowhead marks detail of a clathrin-coated pit
7. Enzyme solution: 1 % (w/v) cellulase (Serva), 1 % (w/v)
pectolyase Y-23 (Kyowa Chemical Products Co. LTD, Tokyo,
Japan), and 0.45 M mannitol (Sigma-Aldrich; see Note 1 ).
8. Wash buffer: 10 mM PIPES (Sigma-Aldrich), pH 6.4, 100 mM
KCl (VWR International Ltd., Lutterworth, UK), and 0.285
mannitol (Sigma-Aldrich; see Note 2 ).
9. Lysis buffer: 10 mM Tris-HCl (Melford Laboratories Ltd.,
Ipswich, UK), pH 7.2, 5 mM MgCl 2 (Sigma-Aldrich), 2 mM