Imaging Plant Nuclei and Membrane-Associated Cytoskeleton by Field Emission Scanning Electron Microscopy - Plant Cell Morphogenesis: Methods and Protocols - page 168

Fig. 1 Tobacco BY-2 cell nucleus and details of a nuclear envelope viewed by feiSEM. ( a ) Nucleus. ( b ) Detail of
the nuclear envelope viewed from a cytoplasmic side. Arrows point the nuclear pore complexes. ( c ) Nuclear
envelope viewed from a nucleoplasmic side after wet fracturing of the nucleus. Remnants of chromatin are
marked by arrow . ( d ) Detail of the nuclear envelope and nuclear pore complex baskets ( encircled ) from a
nucleoplasmic side. Remnants of chromatin are marked by arrow . Remnants of the inner nuclear membrane
are depicted by arrowhead

2 Materials

2.1 Preparing
the Nuclei and a
Disclosure of the
Membrane- Associated
Cytoskeleton

1. Nicotiana tabacum L. cv. Bright Yellow 2 cells.
2. MS medium: 1x MS salts, 1 mg/l thiamine, 200 mg/l
KH 2 PO 4 , 100 mg/l inositol, 30 g/l sucrose, and 0.2 mg/l
2,4-dichlorophenoxyacetic acid (2,4-D), pH 5.8 (all chemi-
cals obtained from Sigma-Aldrich, St. Louis, USA).
3. 100 ml Erlenmeyer fl asks.
4. Horizontal shaker with orbital diameter 30 mm (IKA KS501,
IKA Labortechnik, Germany).
5. 50 ml Falcon tubes.
6. Centrifuge (Sigma 3 K18, Rotor Nr. 11133, 47/100, Sigma
Laboratory Centrifuges, Osterode am Harz, Germany).