Tween 20 solution

The dirt observed to have blocked the surface of the samples was washed off so as to reduce the ubiquity. Then, the leaves samples belonging to each developmental stage and the stem sections were washed with distilled water and, subsequently, with few drops of Bio-Rad tween-20 solution. The leaves and stem sections were washed once with distilled water. The sections were sectioned and fixed using glutaraldehyde (2.5%) in phosphate buffer (0.1 M, pH 7.2) for 24 h. The samples were preserved and made stable by allowing buffer washes thrice for 5 min and fixed in osmium tetroxide (0.5%) for 2 h in the absence of sunlight. Samples were later dehydrated in serial solutions (i.e., 25%, 50%, 70% and 100%). The Quorum K180 critical point dryer was used to dry the samples and later placed on aluminium stubs with the aid of carbon conductive tape and sputter-coated with gold in a QurumQ150 RES gold coater. The samples were viewed with LEO 1450 SEM (SmartSEM) and images were captured and analysed.