适用于扫描电镜的拟南芥蜡质样品制备方法

Comparison of sample preparation methods for scanning electron microscopy( SEM) of leaf epicuticular waxes in Arabidopsis

扫描电镜是观察植物表皮蜡质结构的极佳方法。由于蜡质成分不同,不同的植物有不同的样品干燥方法。本文以拟南芥叶片为实验材料,比较FAA固定和戊二醛-锇酸双固定,CO2临界点干燥和冷冻干燥,以及乙醇、乙酸异戊酯、丙酮和叔丁醇等不同的固定方法、不同的干燥方法和不同的有机溶剂对蜡质的溶解作用后,认为更适于拟南芥表皮蜡质观察的样品制备方法是:戊二醛-锇酸双固定,叔丁醇与乙醇梯度脱水,叔丁醇冷冻干燥。

Waxy layer of the plant epidermis is the first barrier to resistant environmental stimulation. Scanning electron microscopy （ SEM） is a useful technique to observe the leaf epicuticular waxes of Arabidopsis. Different sample preparation methods for SEM observation were compared including critical point drying and freeze drying, FAA fixation and glutaraldehyde-osmium tetroxide double fixation. The effects of different organic solvents on wax structure were also studied. The optimal method to examine epicuticular waxes contains the following processes：glutaraldehyde-osmium tetroxide double fixation, dehydration of tert-butanol and ethanol gradient, and freeze drying.