A New Technique of Preparing Pollen for Scanning Electron Microscopy

A new technique which avoids the distortion usually present in acetolyzed pollen is outlined. The main steps include hydration in Aerosol-OT, ultrasonication in acetone: water, dehydration in ethanol, transfer to amyl acetate, and critical point drying. Experimental studies, in which pollen treated by the new technique is compared with untreated and with acetolyzed pollen, show much better expansion and more observable detail of exine and colpar structures in the pollen prepared by the new method. Damage to Euphorbiaceous grains, especially those with “crotonoid” ornamentation, is extensive using conventional acetolysis but is circumvented entirely by the critical point drying method. It is concluded that SEM and light microscope studies of pollen should include at least some preparations by a non-acetolytic method such as the present one in order to record an optimal amount of structural information.