Scanning Electron Microscopy Preparation of Plant Buds

Buds were fixed in an excess of ice-cold Carnoy (EtOH absolute:Acetic acid glacial 3:1) with the application of a vacuum for 15 min for three times with Carnoy exchanges, and then left overnight in Carnoy in the cold room under weak stirring. The next day, Carnoy was replaced by ice-cold 70–75% EtOH and the samples stored at 4 °C until use. Before scanning, samples were dissected by using a light microscope, while emerging in 70% EtOH, then two times washed in 96% EtOH, each for 20 min., and two times in 96% acetone, each for 30 min. Tissue drying, mounting, and coating followed Pramanik et al. [84 (link)]. Samples were observed with a field emission scanning electron microscope (JSM-7600F SEM, JEOL Ltd., Tokyo, Japan) using the settings and picture sizes described [84 (link)].