Figure 2

A simple method for the isolation of salt glands from A . officinalis. (A) The lower epidermal layer (*) of the leaf was removed with a razor blade to reveal the mesophyll-palisade cell layers (**) prior to enzyme digestion. Bar = 1 cm. (B, C) The leaf with the lower epidermal layer removed were cut into strips and incubated in enzyme solution (B) to obtain the adaxial epidermal peels (C). The epidermal peels appear green (C; left image) due to the presence of mesophyll-palisade cells underneath the epidermal layer, which can be easily removed to obtain peels devoid of chlorophyll-containing cells (C; right image). Bars = 1 cm. (D-G) Salt glands (arrows) were then released through the grinding of these peels (C; right image) followed by a series of purification processes, which include filtering through a 100 μm (pore size) cell strainer (D). The isolated salt glands were finally collected on a 20 μm (pore size) cup filcon (E). All isolated salt glands (D-G) were observed under Olympus inverted light microscope (IMT-2), with close-up observations of purified salt glands viewed under different magnifications (F, G). Bars in D and E = 200 μm, bar in (F) = 80 μm, and bar in (G) = 40 μm.