content and resuspend salts potentially sedimented on the
bottom. In case of bacterial/fungal contamination immediately
discard it. Add the phytohormones when you start using a new
liquid medium bottle. When you open the last bottle, remember to prepare new culture medium as soon as possible, in order
not to run out of medium when you urgently need it.
2. Add the agar only after the pH has been adjusted and the
medium (brought to the final volume) has been transferred
into the bottle, because agar does not dissolve at room temperature. Use two 500 mL bottles instead of 1 L bottle, because
microwave ovens (used later to melt the medium) are usually
not high enough to host 1 L bottle.
3. Loosen the lid of the bottle before inserting it in the microwave
oven, so that the hot air inside the bottle is not under excessive
Fig. 4 Arabidopsis photosynthetic cell suspension cultures stably expressing a YFP-tagged cytosolic protein
(aequorin) [8] (a, b) and freshly isolated protoplasts from them derived (c, d). Confocal microscopy analyses
were carried out with excitation at 488 nm, emission at 505–530 nm for YFP and at 680–720 nm for
chlorophyll (highlighting chloroplasts in red). An overlay of the two channels is shown. Bars: (a–c) 50 μm;
(b, d) 10 μm
Cell Suspension Cultures
181
bottom. In case of bacterial/fungal contamination immediately
discard it. Add the phytohormones when you start using a new
liquid medium bottle. When you open the last bottle, remember to prepare new culture medium as soon as possible, in order
not to run out of medium when you urgently need it.
2. Add the agar only after the pH has been adjusted and the
medium (brought to the final volume) has been transferred
into the bottle, because agar does not dissolve at room temperature. Use two 500 mL bottles instead of 1 L bottle, because
microwave ovens (used later to melt the medium) are usually
not high enough to host 1 L bottle.
3. Loosen the lid of the bottle before inserting it in the microwave
oven, so that the hot air inside the bottle is not under excessive
Fig. 4 Arabidopsis photosynthetic cell suspension cultures stably expressing a YFP-tagged cytosolic protein
(aequorin) [8] (a, b) and freshly isolated protoplasts from them derived (c, d). Confocal microscopy analyses
were carried out with excitation at 488 nm, emission at 505–530 nm for YFP and at 680–720 nm for
chlorophyll (highlighting chloroplasts in red). An overlay of the two channels is shown. Bars: (a–c) 50 μm;
(b, d) 10 μm
Cell Suspension Cultures
181